Du H

References (63)

Title : Interactions between forsythoside E and two cholinesterases at the different conditions: Fluorescence sections - Lin_2024_Methods.Appl.Fluoresc__
Author(s) : Lin C , Du H
Ref : Methods Appl Fluoresc , : , 2024
Abstract : Forsythoside E is one secondary metabolite of Forsythia suspensa (Thunb.) Vahl. In the study, the interactions between forsythoside E and two types of cholinesterases, acetylcholinesterase and butyrylcholinesterase were investigated in the different conditions. Forsythoside E increased the fluorescence intensity of acetylcholinesterase but quenched the fluorescence of butyrylcholinesterase. Abeta25-35 used in the study may not form complexes with cholinesterases, and did not affect the interaction between forsythoside E and cholinesterases. The charged quaternary group of AsCh interacted with the 'anionic' subsite in acetylcholinesterase, which did not affect the interaction between forsythoside E and acetylcholinesterase. The enhancement rate of forsythoside E to acetylcholinesterase fluorescence from high to low was acid solution (pH 6.4), neutral solution (pH 7.4) and alkaline solution (pH 8.0), while the reduction rate of forsythoside E to butyrylcholinesterase fluorescence was in reverse order. Metal ions may interact with cholinesterases, and increased the effects of forsythoside E to cholinesterases fluorescence, in order that Fe3+ was the highest, followed by Cu2+, and Mg2+. A forsythoside E-butyrylcholinesterase complex at stoichiometric ratio of 1:1 was spontaneously formed, and the static quenching was the main quenching mode in the process of forsythoside E binding with butyrylcholinesterase. The K values of two complexes were pretty much the same, suggesting that the interaction between cholinesterases and forsythoside E was almost unaffected by acid-base environment and metal ions. The n numbers of two cholinesterases approximately equaled to one, indicating that there was only one site on each cholinesterase applicable for forsythoside E to bind to.
ESTHER : Lin_2024_Methods.Appl.Fluoresc__
PubMedSearch : Lin_2024_Methods.Appl.Fluoresc__
PubMedID: 38428023

Title : Characterization of lysosomal acid lipase in Ly6G(+) and CD11c(+) myeloid-derived suppressor cells - Zhao_2024_Methods.Cell.Biol_184_119
Author(s) : Zhao T , Du H , Yan C
Ref : Methods Cell Biol , 184 :119 , 2024
Abstract : Lysosomal acid lipase (LAL) is a key enzyme in the metabolic pathway of neutral lipids, whose deficiency (LAL-D) induces the differentiation of myeloid lineage cells into myeloid-derived suppressor cells (MDSCs), which promotes tumor growth and metastasis. This protocol provides detailed procedures for assessment of various LAL biochemical and physiological activities in Ly6G(+) and CD11c(+) MDSCs, including isolation of Ly6G(+) and CD11c(+) cells from the bone marrow and blood of mice, assays of LAL-D-induced cellular metabolic and mitochondrial activities, assessment of LAL-D-induced pathogenic immunosuppressive activity and tumor stimulatory activity. Pharmacological inhibition of the LAL activity was also described in both murine myeloid cells and human white blood cells.
ESTHER : Zhao_2024_Methods.Cell.Biol_184_119
PubMedSearch : Zhao_2024_Methods.Cell.Biol_184_119
PubMedID: 38555152
Gene_locus related to this paper: human-LIPA

Title : ANGPTL4 May Regulate the Crosstalk Between Intervertebral Disc Degeneration and Type 2 Diabetes Mellitus: A Combined Analysis of Bioinformatics and Rat Models - Chen_2023_J.Inflamm.Res_16_6361
Author(s) : Chen Y , Du H , Wang X , Li B , Chen X , Yang X , Zhao C , Zhao J
Ref : J Inflamm Res , 16 :6361 , 2023
Abstract : INTRODUCTION: The crosstalk between intervertebral disc degeneration (IVDD) and type 2 diabetes mellitus (T2DM) has been investigated. However, the common mechanism underlying this phenomenon has not been clearly elucidated. This study aimed to explore the shared gene signatures of IVDD and T2DM. METHODS: The expression profiles of IVDD (GSE27494) and T2DM (GSE20966) were acquired from the Gene Expression Omnibus database. Five hub genes including ANGPTL4, CCL2, CCN3, THBS2, and INHBA were preliminarily screened. GO (Gene Ontology) enrichment analysis, functional correlation analysis, immune filtration, Transcription factors (TFs)-mRNA-miRNA coregulatory network, and potential drugs prediction were performed following the identification of hub genes. RNA sequencing, in vivo and in vitro experiments on rats were further performed to validate the expression and function of the target gene. RESULTS: Five hub genes (ANGPTL4, CCL2, CCN3, THBS2, and INHBA) were identified. GO analysis demonstrated the regulation of the immune system, extracellular matrix (ECM), and SMAD protein signal transduction. There was a strong correlation between hub genes and different functions, including lipid metabolism, mitochondrial function, and ECM degradation. The immune filtration pattern grouped by disease and the expression of hub genes showed significant changes in the immune cell composition. TFs-mRNA-miRNA co-expression networks were constructed. In addition, pepstatin showed great drug-targeting relevance based on potential drugs prediction of hub genes. ANGPTL4, a gene that mediates the inhibition of lipoprotein lipase activity, was eventually determined after hub gene screening, validation by different datasets, RNA sequencing, and experiments. DISCUSSION: This study screened five hub genes and ANGPTL4 was eventually determined as a potential target for the regulation of the crosstalk in patients with IVDD and T2DM.
ESTHER : Chen_2023_J.Inflamm.Res_16_6361
PubMedSearch : Chen_2023_J.Inflamm.Res_16_6361
PubMedID: 38161353

Title : Novel harmine derivatives as potent acetylcholinesterase and amyloid beta aggregation dual inhibitors for management of Alzheimer's disease - Du_2023_J.Enzyme.Inhib.Med.Chem_38_2281893
Author(s) : Du H , Song J , Ma F , Gao H , Zhao X , Mao R , He X , Yan Y
Ref : J Enzyme Inhib Med Chem , 38 :2281893 , 2023
Abstract : In this study, a series of potential ligands for the treatment of AD were synthesised and characterised as novel harmine derivatives modified at position 9 with benzyl piperazinyl. In vitro studies revealed that the majority of the derivatives exhibited moderate to potent inhibition against hAChE and Abeta(1-42) aggregation. Notably, compounds 13 and 17d displayed potent drug-likeness and ADMET properties, demonstrating remarkable inhibitory activities towards AChE (IC(50) = 58.76 nM and 89.38 nM, respectively) as well as Abeta aggregation (IC(50) = 9.31 microM and 13.82 microM, respectively). More importantly, compounds 13 and 17d showed exceptional neuroprotective effects against Abeta(1-42)-induced SH-SY5Y damage, while maintaining low toxicity in SH-SY5Y cells. Further exploration of the mechanism through kinetic studies and molecular modelling confirmed that compound 13 could interact with both the CAS and the PAS of AChE. These findings suggested that harmine derivatives hold great potential as dual-targeted candidates for treating AD.
ESTHER : Du_2023_J.Enzyme.Inhib.Med.Chem_38_2281893
PubMedSearch : Du_2023_J.Enzyme.Inhib.Med.Chem_38_2281893
PubMedID: 37965884

Title : Substrate-enzyme interactions and catalytic mechanism in a novel family VI esterase with dibutyl phthalate-hydrolyzing activity - Cheng_2023_Environ.Int_178_108054
Author(s) : Cheng J , Du H , Zhou MS , Ji Y , Xie YQ , Huang HB , Zhang SH , Li F , Xiang L , Cai QY , Li YW , Li H , Li M , Zhao HM , Mo CH
Ref : Environ Int , 178 :108054 , 2023
Abstract : Microbial degradation has been confirmed as effective and environmentally friendly approach to remediate phthalates from the environment, and hydrolase is an effective element for contaminant degradation. In the present study, a novel dibutyl phthalate (DBP)-hydrolyzing carboxylesterase (named PS06828) from Pseudomonas sp. PS1 was heterogeneously expressed in E. coli, which was identified as a new member of the lipolytic family VI. Purified PS06828 could efficiently degrade DBP with a wide range of temperature (25-37 degreesC) and pH (6.5-9.0). Multi-spectroscopy methods combined with molecular docking were employed to study the interaction of PS06828 with DBP. Fluorescence and UV-visible absorption spectra revealed the simultaneous presence of static and dynamic component in the fluorescence quenching of PS06828 by DBP. Synchronous fluorescence and circular dichroism spectra showed inconspicuous alteration in micro-environmental polarity around amino acid residues but obvious increasing of alpha-helix and reducing of beta-sheet and random coil in protein conformation. Based on the information on exact binding sites of DBP on PS06828 provided by molecular docking, the catalytic mechanism mediated by key residues (Ser113, Asp166, and His197) was proposed and subsequently confirmed by site-directed mutagenesis. The results can strengthen our mechanistic understanding of family VI esterase involved in hydrolysis of phthalic acid esters, and provide a solid foundation for further enzymatic modification.
ESTHER : Cheng_2023_Environ.Int_178_108054
PubMedSearch : Cheng_2023_Environ.Int_178_108054
PubMedID: 37354883
Gene_locus related to this paper: pseae-PA3859

Title : LAL deficiency induced myeloid-derived suppressor cells as targets and biomarkers for lung cancer - Zhao_2023_J.Immunother.Cancer_11_
Author(s) : Zhao T , Liu S , Hanna NH , Jalal S , Ding X , Wan J , Yan C , Du H
Ref : J Immunother Cancer , 11 : , 2023
Abstract : BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells in tumor microenvironment, which suppress antitumor immunity. Expansion of various MDSC subpopulations is closely associated with poor clinical outcomes in cancer. Lysosomal acid lipase (LAL) is a key enzyme in the metabolic pathway of neutral lipids, whose deficiency (LAL-D) in mice induces the differentiation of myeloid lineage cells into MDSCs. These Lal (-/-) MDSCs not only suppress immune surveillance but also stimulate cancer cell proliferation and invasion. Understanding and elucidating the underlying mechanisms of MDSCs biogenesis will help to facilitate diagnosis/prognosis of cancer occurrence and prevent cancer growth and spreading. METHODS: Single-cell RNA sequencing (scRNA-seq) was performed to distinguish intrinsic molecular and cellular differences between normal versus Lal (-/-) bone marrow-derived Ly6G(+) myeloid populations in mice. In humans, LAL expression and metabolic pathways in various myeloid subsets of blood samples of patients with non-small cell lung cancer (NSCLC) were assessed by flow cytometry. The profiles of myeloid subsets were compared in patients with NSCLC before and after the treatment of programmed death-1 (PD-1) immunotherapy. RESULTS: scRNA-seq of Lal (-/-) CD11b(+)Ly6G(+) MDSCs identified two distinctive clusters with differential gene expression patterns and revealed a major metabolic shift towards glucose utilization and reactive oxygen species (ROS) overproduction. Blocking pyruvate dehydrogenase (PDH) in glycolysis reversed Lal (-/-) MDSCs' capabilities of immunosuppression and tumor growth stimulation and reduced ROS overproduction. In the blood samples of human patients with NSCLC, LAL expression was significantly decreased in CD13(+)/CD14(+)/CD15(+)/CD33(+) myeloid cell subsets. Further analysis in the blood of patients with NSCLC revealed an expansion of CD13(+)/CD14(+)/CD15(+) myeloid cell subsets, accompanied by upregulation of glucose-related and glutamine-related metabolic enzymes. Pharmacological inhibition of the LAL activity in the blood cells of healthy participants increased the numbers of CD13(+) and CD14(+) myeloid cell subsets. PD-1 checkpoint inhibitor treatment in patients with NSCLC reversed the increased number of CD13(+) and CD14(+) myeloid cell subsets and PDH levels in CD13(+) myeloid cells. CONCLUSION: These results demonstrate that LAL and the associated expansion of MDSCs could serve as targets and biomarkers for anticancer immunotherapy in humans.
ESTHER : Zhao_2023_J.Immunother.Cancer_11_
PubMedSearch : Zhao_2023_J.Immunother.Cancer_11_
PubMedID: 36914206
Gene_locus related to this paper: mouse-1llip

Title : Therapeutic efficacy of rscAAVrh74.miniCMV.LIPA gene therapy in a mouse model of lysosomal acid lipase deficiency - Lam_2022_Mol.Ther.Methods.Clin.Dev_26_413
Author(s) : Lam P , Ashbrook A , Zygmunt DA , Yan C , Du H , Martin PT
Ref : Mol Ther Methods Clin Dev , 26 :413 , 2022
Abstract : Lysosomal acid lipase deficiency (LAL-D) presents as one of two rare autosomal recessive diseases: Wolman disease (WD), a severe disorder presenting in infancy characterized by absent or very low LAL activity, and cholesteryl ester storage disease (CESD), a less severe, later onset disease form. Recent clinical studies have shown efficacy of enzyme replacement therapy for both forms of LAL-D; however, no gene therapy approach has yet been developed for clinical use. Here, we show that rscAAVrh74.miniCMV.LIPA gene therapy can significantly improve disease symptoms in the Lipa (-/-) mouse model of LAL-D. Treatment dramatically lowered hepatosplenomegaly, liver and spleen triglyceride and cholesterol levels, and serum expression of markers of liver damage. Measures of liver inflammation and fibrosis were also reduced. Treatment of young adult mice was more effective than treatment of neonates, and enzyme activity was elevated in serum, consistent with possible bystander effects. These results demonstrate that adeno associated virus (AAV)-mediated LIPA gene-replacement therapy may be a viable option to treat patients with LAL-D, particularly patients with CESD.
ESTHER : Lam_2022_Mol.Ther.Methods.Clin.Dev_26_413
PubMedSearch : Lam_2022_Mol.Ther.Methods.Clin.Dev_26_413
PubMedID: 36092360
Gene_locus related to this paper: mouse-1llip , human-LIPA

Title : Identification of closely associated SNPs and candidate genes with seed size and shape via deep re-sequencing GWAS in soybean - Shao_2022_Theor.Appl.Genet__
Author(s) : Shao Z , Shao J , Huo X , Li W , Kong Y , Du H , Li X , Zhang C
Ref : Theor Appl Genet , : , 2022
Abstract : A soybean natural population was genotyped by deep re-sequencing and phenotyped for six seed size- and shape-related traits under six environments to identify closely associated SNPs and candidate genes. Seed size and shape are important determining factors for soybean yield formation, while their genetic basis and molecular mechanism are still largely unknown, which seriously constrains the increasing of soybean yield at present. In view of this, a natural population was genotyped via the deep re-sequencing technique (~ 20 x) and phenotyped for six related traits under six environments. In total, 154 SNPs were closely associated with seed length across diverse environments, and 323, 483, 565, 394 and 2038 SNPs were closely associated with seed width, seed diameter, seed circumference, seed area and ratio of length to width under multiple environments. Moreover, 98.70%, 96.28%, 48.24%, 85.13%, 97.21% and 98.58% of them were further demonstrated by the BLUP and mean values of the related traits. Furthermore, 218 genes flanking the associated SNPs on chromosomes 6 and 10 were analyzed for DNA mutations and RNA expressions through SNP alleles and transcriptome data, simultaneously. The candidate genes, Glyma.10G035200 (Sn1-specific diacylglycerol lipase), Glyma.10G035400 (transcription factor) and Glyma.10G058200 (phenylalanine ammonia-lyase), were discovered to relate with the seed size and shape for their different DNA sequences or differential RNA expressions among soybean varieties at five seed developmental stages. Thus, these closely associated SNPs and related genes provide novel insights and useful information for the seed size and shape genetic basis dissection and breeding improvement in soybean.
ESTHER : Shao_2022_Theor.Appl.Genet__
PubMedSearch : Shao_2022_Theor.Appl.Genet__
PubMedID: 35588015

Title : The Interaction Between Two Metabolites of Polygala Tenuifolia and Cholinesterases - Gao_2022_Protein.Pept.Lett__
Author(s) : Gao C , Du H
Ref : Protein Pept Lett , : , 2022
Abstract : OBJECTIVE: The work aimed to compare the binding between the two main components of Polygala tenuifolia Willd. and two cholinesterases (ChEs) by using a variety of spectral techniques. METHODS: Two main components of Polygala tenuifolia Willd. included Tenuifolin (Ten) and Onjisaponin B (Onj B), and two cholinesterases (ChEs) included acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). METHODS: The UV-visible absorption spectra results showed that Ten had no effect on the structure of ChEs, and the combination of Onj B with ChEs changed its structure. Onj B statically quenched the endogenous fluorescence of both of ChEs, Ten dynamically quenched the endogenous fluorescence of AChE with no effect on BChE. The fluorescence quenching rate of ChEs by Onj B was much higher than that of AChE by Ten, and only one binding site of each protein spontaneously interacted with the compound to bind to or collide. Synchronous fluorescence results showed that Ten and Onj B quenched the fluorescence intensity by affecting tryptophan and tyrosine residues in cholinesterases, respectively. Hydrophobic force played an important role in the interaction between Ten and AChE, and van der Waals force and hydrogen bond were the main driving forces for the binding of Onj B to ChEs. RESULTS: The Enzyme activity test results showed that Onj B inhibited ChE activity, and Ten never inhibited ChE activity. CONCLUSION: Onj B has the potential to inhibit ChE activity and increase the neurotransmitter acetylcholine content in the nerve system, inproving the Alzheimer's disease (AD).
ESTHER : Gao_2022_Protein.Pept.Lett__
PubMedSearch : Gao_2022_Protein.Pept.Lett__
PubMedID: 36028966

Title : Lysosomal acid lipase, CSF1R and PD-L1 determine functions of CD11c+ myeloid-derived suppressor cells - Zhao_2022_JCI.Insight__
Author(s) : Zhao T , Liu S , Ding X , Johnson EM , Hanna NH , Singh K , Sen CK , Wan J , Du H , Yan C
Ref : JCI Insight , : , 2022
Abstract : Lysosomal acid lipase (LAL) is a key enzyme in the metabolic pathway of neutral lipids. In the blood of LAL deficient (lal-/-) mice, increased CD11c+ cells were accompanied by up-regulated PD-L1 expression. Single cell RNA sequencing of lal-/- CD11c+ cells identified two distinctive clusters with a major metabolic shift towards glucose utilization and reactive oxygen species (ROS) over-production. Pharmacologically blocking pyruvate dehydrogenase in glycolysis not only reduced CD11c+ cells and their PD-L1 expression, but also reversed their capabilities of T cell suppression and tumor growth stimulation. Colony-stimulating factor 1 receptor (CSF1R) plays an essential role in controlling lal-/- CD11c+ cell homeostasis and function and PD-L1 expression. Inhibition of LAL activity by pharmacological inhibitor increased CD11c, PD-L1 and CSF1R levels in both normal murine myeloid cells and human blood cells. Tumor-bearing mice and human non-small-cell lung cancer (NSCLC) patients also showed CD11c+ cell expansion with PD-L1 and CSF1R up-regulation and immunosuppression. There were positive correlations among CD11c, PD-L1 and CSF1R expression and negative correlations with LAL expression in lung cancer and melanoma patients using the TCGA database and patient samples. Therefore, CD11c+ cells switched their functions to immune suppression and tumor growth stimulation through CSF1R/PD-L1 upregulation and metabolic reprogramming.
ESTHER : Zhao_2022_JCI.Insight__
PubMedSearch : Zhao_2022_JCI.Insight__
PubMedID: 35917184

Title : Analysis of the binding selectivity and inhibiting mechanism of chlorogenic acid isomers and their interaction with grass carp endogenous lipase using multi-spectroscopic, inhibition kinetics and modeling methods - Xu_2022_Food.Chem_382_132106
Author(s) : Xu Z , Cao Q , Manyande A , Xiong S , Du H
Ref : Food Chem , 382 :132106 , 2022
Abstract : Polyphenols are inhibitors for lipase, but the binding selectivity and mechanism of polyphenol isomers and how they interact with lipase are not clear. Here, chlorogenic acid (CGA) isomers, neochlorogenic acid (NCGA) and cryptochlorogenic acid (CCGA) were used to explore the binding selectivity and mechanism of lipase. An inhibition assay indicated that both CGA isomers had dose-dependent inhibitory effects on lipase; however, the inhibitory effect of NCGA was better (IC(50): 0.647 mg/mL) than that of CCGA (IC(50): 0.677 mg/mL). NCGA and CCGA formed complexes with lipase at a molar ratio of 1:1, and the electrostatic interaction force plays a major role in the lipase-CCGA system. Molecular dynamics studies demonstrated that NCGA had a greater impact on the structure of lipase. The multi-spectroscopic and modeling results explained the effects of micro-structural changes on the binding site, the interaction force and the inhibition rate of the isomers when they combined with lipase.
ESTHER : Xu_2022_Food.Chem_382_132106
PubMedSearch : Xu_2022_Food.Chem_382_132106
PubMedID: 35240531

Title : Lysosomal Acid Lipase Deficiency Controls T- and B-Regulatory Cell Homeostasis in the Lymph Nodes of Mice with Human Cancer Xenotransplants - Ding_2021_Am.J.Pathol_191_353
Author(s) : Ding X , Zhao T , Lee CC , Yan C , Du H
Ref : American Journal of Pathology , 191 :353 , 2021
Abstract : Utilization of proper preclinical models accelerates development of immunotherapeutics and the study of the interplay between human malignant cells and immune cells. Lysosomal acid lipase (LAL) is a critical lipid hydrolase that generates free fatty acids and cholesterol. Ablation of LAL suppresses immune rejection and allows growth of human lung cancer cells in lal(-/-) mice. In the lal(-/-) lymph nodes, the percentages of both T- and B-regulatory cells (Tregs and Bregs, respectively) are increased, with elevated expression of programmed death-ligand 1 and IL-10, and decreased expression of interferon-gamma. Levels of enzymes in the glucose and glutamine metabolic pathways are elevated in Tregs and Bregs of the lal(-/-) lymph nodes. Pharmacologic inhibitor of pyruvate dehydrogenase, which controls the transition from glycolysis to the citric acid cycle, effectively reduces Treg and Breg elevation in the lal(-/-) lymph nodes. Blocking the mammalian target of rapamycin or reactivating peroxisome proliferator-activated receptor gamma, an LAL downstream effector, reduces lal(-/-) Treg and Breg elevation and PD-L1 expression in lal(-/-) Tregs and Bregs, and improves human cancer cell rejection. Treatment with PD-L1 antibody also reduces Treg and Breg elevation in the lal(-/-) lymph nodes and improves human cancer cell rejection. These observations conclude that LAL-regulated lipid metabolism is essential to maintain antitumor immunity.
ESTHER : Ding_2021_Am.J.Pathol_191_353
PubMedSearch : Ding_2021_Am.J.Pathol_191_353
PubMedID: 33159889

Title : Novel deoxyvasicinone and tetrahydro-beta-carboline hybrids as inhibitors of acetylcholinesterase and amyloid beta aggregation - Du_2020_Bioorg.Med.Chem.Lett__127659
Author(s) : Du H , Jiang X , Ma M , Xu H , Liu S , Ma F
Ref : Bioorganic & Medicinal Chemistry Lett , :127659 , 2020
Abstract : A novel series of deoxyvasicinone-tetrahydro-beta-carboline hybrids were synthesized and evaluated as acetylcholinesterase (AChE) and beta-amyloid peptide (Abeta) aggregation inhibitors for the treatment of Alzheimer's disease. The results revealed that the derivatives had multifunctional profiles, including AChE inhibition, Abeta(1-42) aggregation inhibition, and neuroprotective properties. Inspiringly, hybrids 8b and 8d displayed excellent inhibitory activities against hAChE (IC(50) = 0.93 and 1.08 nM, respectively) and Abeta(1-42) self-aggregation (IC(50) = 19.71 and 2.05 M, respectively). In addition, 8b and 8d showed low cytotoxicity and good neuroprotective activity against Abeta(1-42)-induced damage in SH-SY5Y cells.
ESTHER : Du_2020_Bioorg.Med.Chem.Lett__127659
PubMedSearch : Du_2020_Bioorg.Med.Chem.Lett__127659
PubMedID: 33137375

Title : Mechanistic insight into esterase-catalyzed hydrolysis of phthalate esters (PAEs) based on integrated multi-spectroscopic analyses and docking simulation - Du_2020_J.Hazard.Mater_408_124901
Author(s) : Du H , Hu RW , Zhao HM , Huang HB , Xiang L , Liu BL , Feng NX , Li H , Li YW , Cai QY , Mo CH
Ref : J Hazard Mater , 408 :124901 , 2020
Abstract : A novel PAE-hydrolyzing esterase (named Hyd) gene was screened from the genomic library of Rhodococcus sp. 2G and was successfully expressed in heterologous E. coli, which was defined as a new family of esterolytic enzymes. The purified Hyd could efficiently degrade various PAEs, displaying high activity and stability with a broad range of pH (4-10) and temperature (20-60 degreesC). Interaction mechanism of Hyd with dibutyl phthalate (DBP) was investigated by integrated multi-spectroscopic and docking simulation methods. Fluorescence and UV-vis spectra revealed that DBP could quench the fluorescence of Hyd through a static quenching mechanism. The results from synchronous fluorescence and CD spectra confirmed that the DBP binding to Hyd triggered conformational and micro-environmental changes of Hyd, which were characterized by increased stretching extent and random coil, and decreased alpha-helix and beta-sheet. Molecular docking study showed that DBP could be bound to the cavity of Hyd with hydrogen bonding and hydrophobic interaction. A novel and distinctive catalytic mechanism was proposed: two key residues Thr(190) and Ser(191) might catalyze the hydrolysis of DBP, instead of the conserved catalytic triad (Ser-His-Asp) reported elsewhere, which was confirmed by site-directed mutagenesis.
ESTHER : Du_2020_J.Hazard.Mater_408_124901
PubMedSearch : Du_2020_J.Hazard.Mater_408_124901
PubMedID: 33360702
Gene_locus related to this paper: rhoso-a0a3g5hne7

Title : Lysosomal Acid Lipase Is Required for Donor T Cells to Induce Graft-versus-Host Disease - Nguyen_2020_Cell.Rep_33_108316
Author(s) : Nguyen HD , Ticer T , Bastian D , Kuril S , Li H , Du H , Yan C , Yu XZ
Ref : Cell Rep , 33 :108316 , 2020
Abstract : Graft-versus-host disease (GVHD) limits the success of allogeneic hematopoietic cell transplantation (allo-HCT). Lysosomal acid lipase (LAL) mediates the intrinsic lipolysis of cells to generate free fatty acids (FFAs), which play an essential role in the development, proliferation, and function of T cells. Here, we find that LAL is essential for donor T cells to induce GVHD in murine models of allo-HCT. Specifically, LAL is required for donor T cell survival, differentiation, and alloreactivity in GVHD target organs, but not in lymphoid organs. LAL induces the differentiation of donor T cells toward GVHD pathogenic Th1/Tc1 and Th17 while suppressing regulatory T cell generation. LAL(-/-) T cells succumb to oxidative stress and become anergic in target organs. Pharmacologically targeting LAL effectively prevents GVHD development while preserving the GVL activity. Thus, the present study reveals the role of LAL in T cell alloresponse and pathogenicity and validates LAL as a target for controlling GVHD and tumor relapse after allo-HCT.
ESTHER : Nguyen_2020_Cell.Rep_33_108316
PubMedSearch : Nguyen_2020_Cell.Rep_33_108316
PubMedID: 33113360

Title : Bioaugmentation of Exogenous Strain Rhodococcus sp. 2G Can Efficiently Mitigate Di(2-ethylhexyl) Phthalate Contamination to Vegetable Cultivation - Zhao_2019_J.Agric.Food.Chem_67_6940
Author(s) : Zhao HM , Du H , Huang CQ , Li S , Zeng XH , Huang XJ , Xiang L , Li H , Li YW , Cai QY , Mo CH , He Z
Ref : Journal of Agricultural and Food Chemistry , 67 :6940 , 2019
Abstract : This work developed a bioaugmentation strategy that simultaneously reduced soil di(2-ethylhexyl) phthalate (DEHP) pollution and its bioaccumulation in Brassica parachinensis by inoculating the isolated strain Rhodococcus sp. 2G. This strain could efficiently degrade DEHP at a wide concentration range from 50 to 1600 mg/L and transformed DEHP through a unique biochemical degradation pathway that distinguished it from other Rhodococcus species. Besides, strain 2G colonized well in the rhizosphere soil of the inoculated vegetable without competition with indigenous microbes, resulting in increased removal of DEHP from soil (95%) and reduced DEHP bioaccumulation in vegetables (75% in the edible part) synchronously. Improved enzyme activities and DOC content in the rhizosphere of the planting vegetable and inoculating strain 2G were responsible for the high efficiency in mitigating DEHP contamination to vegetable cultivation. This work demonstrated a great potential application to grow vegetables in contaminated soil for safe food production.
ESTHER : Zhao_2019_J.Agric.Food.Chem_67_6940
PubMedSearch : Zhao_2019_J.Agric.Food.Chem_67_6940
PubMedID: 31021627
Gene_locus related to this paper: rhoso-a0a3g5hne7

Title : Engineering Pseudomonas entomophila for synthesis of copolymers with defined fractions of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates - Li_2019_Metab.Eng_52_253
Author(s) : Li M , Chen X , Che X , Zhang H , Wu LP , Du H , Chen GQ
Ref : Metab Eng , 52 :253 , 2019
Abstract : Polyhydroxyalkanoates (PHA) composed of both short-chain-length (SCL) and medium-chain-length (MCL) monomers (SCL-co-MCL PHA) combine the advantages of high strength and elasticity provided by SCL PHA and MCL PHA, respectively. Synthesis of SCL-co-MCL PHA, namely, copolymers of 3-hydroxybutyrate (3HB) and MCL 3-hydroxyalkanoates (3HA) such as 3-hydroxydecanoate (3HD) and longer chain 3HA, has been a challenge for a long time. This study aims to engineer Pseudomonas entomophila for synthesizing P(3HB-co-MCL 3HA) via weakening its beta-oxidation pathway combined with insertion of 3HB synthesis pathway consisting of beta-ketothiolase (phaA) and acetoacetyl-CoA reductase (phaB). 3HB and MCL 3HA polymerization is catalyzed by a low specificity PHA synthase (phaC), namely, mutated PhaC61-3. The link between the fatty acid de novo synthesis and PHA synthesis was further blocked to increase the supply for SCL and MCL monomers in P. entomophila. The so-constructed P. entomophila was successfully used to synthesize novel PHA copolymers of P(3HB-co-3HD), P(3HB-co-3HDD) and P(3HB-co-3H9D) consisting of 3HB and 3-hydroxydecanoate (3HD), 3-hydroxydodecanoate (3HDD) and 3-hydroxy-9-decanent (3H9D), respectively. MCL 3HA compositions of P(3HB-co-3HD) and P(3HB-co-3HDD) can be adjusted from 0 to approximate 100 mol%. Results demonstrated that the engineered P. entomophila could be a platform for tailor-made P(3HB-co-MCL 3HA).
ESTHER : Li_2019_Metab.Eng_52_253
PubMedSearch : Li_2019_Metab.Eng_52_253
PubMedID: 30582985

Title : Study of the interactions of forsythiaside and rutin with acetylcholinesterase (AChE) - Yan_2018_Int.J.Biol.Macromol_119_1344
Author(s) : Yan X , Chen T , Zhang L , Du H
Ref : Int J Biol Macromol , 119 :1344 , 2018
Abstract : Acetylcholinesterase (AChE) inhibitors have been considered as candidates for the treatment of Alzheimer's disease (AD) and have been utilized in clinical trials. In the present study, the interactions of forsythiaside and rutin with AChE have been investigated, after discovering the inhibitory AChE activity of the two compounds. Forsythiaside and rutin both can bind to AChE to form forsythiaside-AChE and rutin-AChE complex, and thus quench the intrinsic fluorescence of AChE. The quenching mechanism, the binding sites, the binding forces, the binding constants and the energy transfer involved were studied in details. Forsythiaside and rutin show some properties in common, including the stoichiometric binding ratio of 1:1 with AChE and the full quenching of AChE fluorescence. At the same time, the two compounds distinctly present some different characters, for example, the binding constant of rutin is less than that of forsythiaside, and the interaction force and the affinity between forsythiaside and AChE are much bigger than that of rutin. Spectroscopy data and docking analysis powerfully support the findings that forsythiaside inhibit AChE activity more strongly than rutin. The current study will provide the better understanding on the nature of the possible interactions between forsythiaside and rutin with AChE.
ESTHER : Yan_2018_Int.J.Biol.Macromol_119_1344
PubMedSearch : Yan_2018_Int.J.Biol.Macromol_119_1344
PubMedID: 30048725

Title : Endothelial Rab7 GTPase mediates tumor growth and metastasis in lysosomal acid lipase-deficient mice - Zhao_2017_J.Biol.Chem_292_19198
Author(s) : Zhao T , Ding X , Yan C , Du H
Ref : Journal of Biological Chemistry , 292 :19198 , 2017
Abstract : Tumors depend on their microenvironment for sustained growth, invasion, and metastasis. In this environment, endothelial cells (ECs) are an important stromal cell type interacting with malignant cells to facilitate tumor angiogenesis and cancer cell extravasation. Of note, lysosomal acid lipase (LAL) deficiency facilitates melanoma growth and metastasis. ECs from LAL-deficient (lal(-/-)) mice possess enhanced proliferation, migration, and permeability of inflammatory cells by activating the mammalian target of rapamycin (mTOR) pathway. Here we report that lal(-/-) ECs facilitated in vivo tumor angiogenesis, growth, and metastasis, largely by stimulating tumor cell proliferation, migration, adhesion, and transendothelial migration via increased expression of IL-6 and monocyte chemoattractant protein 1 (MCP-1). This prompted us to look for lysosomal proteins that are involved in lal(-/-) EC dysfunctions. We found that lal(-/-) ECs displayed increased expression of Rab7, a late endosome/lysosome-associated small GTPase. Moreover, Rab7 and mTOR were co-increased and co-localized to lysosomes and physically interacted in lal(-/-) ECs. Rab7 inhibition reversed lal(-/-) EC dysfunctions, including decreasing their enhanced migration and permeability of tumor-stimulatory myeloid cells, and suppressed EC-mediated stimulation of in vitro tumor cell transmigration, proliferation, and migration and in vivo tumor growth and metastasis. Finally, Rab7 inhibition reduced overproduction of reactive oxygen species and increased IL-6 and MCP-1 secretion in lal(-/-) ECs. Our results indicate that metabolic reprogramming resulting from LAL deficiency enhances the ability of ECs to stimulate tumor cell proliferation and metastasis through stimulation of lysosome-anchored Rab7 activity.
ESTHER : Zhao_2017_J.Biol.Chem_292_19198
PubMedSearch : Zhao_2017_J.Biol.Chem_292_19198
PubMedID: 28924047
Gene_locus related to this paper: mouse-1llip

Title : Lysosome-mediated degradation of a distinct pool of lipid droplets during hepatic stellate cell activation - Tuohetahuntila_2017_J.Biol.Chem_292_12436
Author(s) : Tuohetahuntila M , Molenaar MR , Spee B , Brouwers JF , Wubbolts R , Houweling M , Yan C , Du H , VanderVen BC , Vaandrager AB , Helms JB
Ref : Journal of Biological Chemistry , 292 :12436 , 2017
Abstract : Activation of hepatic stellate cells (HSCs) is a critical step in the development of liver fibrosis. During activation, HSCs lose their lipid droplets (LDs) containing triacylglycerols (TAGs), cholesteryl esters, and retinyl esters (REs). We previously provided evidence for the presence of two distinct LD pools, a preexisting and a dynamic LD pool. Here we investigate the mechanisms of neutral lipid metabolism in the preexisting LD pool. To investigate the involvement of lysosomal degradation of neutral lipids, we studied the effect of lalistat, a specific lysosomal acid lipase (LAL/Lipa) inhibitor on LD degradation in HSCs during activation in vitro The LAL inhibitor increased the levels of TAG, cholesteryl ester, and RE in both rat and mouse HSCs. Lalistat was less potent in inhibiting the degradation of newly synthesized TAG species as compared with a more general lipase inhibitor orlistat. Lalistat also induced the presence of RE-containing LDs in an acidic compartment. However, targeted deletion of the Lipa gene in mice decreased the liver levels of RE, most likely as the result of a gradual disappearance of HSCs in livers of Lipa(-/-) mice. Lalistat partially inhibited the induction of activation marker alpha-smooth muscle actin (alpha-SMA) in rat and mouse HSCs. Our data suggest that LAL/Lipa is involved in the degradation of a specific preexisting pool of LDs and that inhibition of this pathway attenuates HSC activation.
ESTHER : Tuohetahuntila_2017_J.Biol.Chem_292_12436
PubMedSearch : Tuohetahuntila_2017_J.Biol.Chem_292_12436
PubMedID: 28615446

Title : Novel deoxyvasicinone derivatives as potent multitarget-directed ligands for the treatment of Alzheimer's disease: Design, synthesis, and biological evaluation - Ma_2017_Eur.J.Med.Chem_140_118
Author(s) : Ma F , Du H
Ref : Eur Journal of Medicinal Chemistry , 140 :118 , 2017
Abstract : A series of multitarget ligands was designed by introducing several structurally diverse aminoacetamide groups at position 6 of the deoxyvasicinone group, with the aim of obtaining novel multifunctional anti-Alzheimer's disease agents using deoxyvasicinone as the substrate. In vitro studies showed that almost all of the derivatives were potent inhibitors of human recombinant acetylcholinesterase (hAChE) and human serum butyrylcholinesterase (hBChE), with IC50 values in the low nanomolar range, and exhibited moderate to high inhibition of Abeta1-42 self-aggregation. In particular, compounds 12h, 12n, and 12q showed promising inhibitory activity for hAChE, with IC50 values of 5.31 +/- 2.8, 4.09 +/- 0.23, and 7.61 +/- 0.53 nM, respectively. Compounds 12h and 12q also exhibited the greatest ability to inhibit hBChE, with IC50 values of 4.35 +/- 0.32 and 2.35 +/- 0.14 nM, respectively. Moreover, enzyme kinetics confirmed that compound 12q caused a mixed type of AChE inhibition, by binding to both the active sites (PAS and CAS) of AChE. Remarkably, compound 12q also demonstrated the highest potential inhibitory activity for Abeta1-42 self-aggregation (63.9 +/- 4.9%, 10 muM), and it was also an excellent metal chelator.
ESTHER : Ma_2017_Eur.J.Med.Chem_140_118
PubMedSearch : Ma_2017_Eur.J.Med.Chem_140_118
PubMedID: 28923380

Title : A new method to characterize the kinetics of cholinesterases inhibited by carbamates - Xiao_2017_J.Pharm.Biomed.Anal_144_175
Author(s) : Xiao Q , Zhou H , Wei H , Du H , Tan W , Zhan Y , Pistolozzi M
Ref : J Pharm Biomed Anal , 144 :175 , 2017
Abstract : The inhibition of cholinesterases (ChEs) by carbamates includes a carbamylation (inhibition) step, in which the drug transfers its carbamate moiety to the active site of the enzyme and a decarbamylation (activity recovery) step, in which the carbamyl group is hydrolyzed from the enzyme. The carbamylation and decarbamylation kinetics decide the extent and the duration of the inhibition, thus the full characterization of candidate carbamate inhibitors requires the measurement of the kinetic constants describing both steps. Carbamylation and decarbamylation rate constants are traditionally measured by two separate set of experiments, thus making the full characterization of candidate inhibitors time-consuming. In this communication we show that by the analysis of the area under the inhibition-time curve of cholinesterases inhibited by carbamates it is possible to calculate the decarbamylation rate constant from the same data traditionally used to characterize only the carbamylation kinetics, therefore it is possible to obtain a full characterization of the inhibition with a single set of experiments. The characterization of the inhibition kinetics of human and dog plasma butyrylcholinesterase and of human acetylcholinesterase by bambuterol and bambuterol monocarbamate enantiomers was used to demonstrate the validity of the approach. The results showed that the proposed method provides reliable estimations of carbamylation and decarbamylation rate constants thus representing a simple and useful approach to reduce the time required for the characterization of carbamate inhibitors.
ESTHER : Xiao_2017_J.Pharm.Biomed.Anal_144_175
PubMedSearch : Xiao_2017_J.Pharm.Biomed.Anal_144_175
PubMedID: 28483282

Title : Protective effects of Forsythoside A on amyloid beta-induced apoptosis in PC12 cells by downregulating acetylcholinesterase - Yan_2017_Eur.J.Pharmacol_810_141
Author(s) : Yan X , Chen T , Zhang L , Du H
Ref : European Journal of Pharmacology , 810 :141 , 2017
Abstract : Increasing the acetylcholine level and fighting the neuroinflammation has always been taken as a treatment strategy for Alzheimer's disease (AD). Forsythoside A is a major component in Forsythia suspensa (Thunb.) Vahl (F. suspensa, Lianqiao in Chinese) that has been traditionally used as Chinese herbal medicine to treat the inflammation in China. This study examined the inhibitory acetylcholinesterase activities of Forsythoside A at chemical and biological level. Forsythoside A inhibited acetylcholinesterase in a mixed type of inhibition, with Ki of 47.68muM. Docking analysis strongly supported these findings. In PC12 cells Forsythoside A increased cell viability and suppressed acetylcholinesterase increased by Abeta25-35, thus alleviated the corresponding apoptosis. Taken together, these results suggest that Forsythoside A has the protective effects on Abeta25-35-induced apoptosis in PC12 cells by downregulating acetylcholinesterase, making it a potential functional food ingredient or drug candidate for the treatment of AD.
ESTHER : Yan_2017_Eur.J.Pharmacol_810_141
PubMedSearch : Yan_2017_Eur.J.Pharmacol_810_141
PubMedID: 28687196

Title : A New Flavonoid Glycoside from Lysionotus pauciflorus - Luo_2016_Nat.Prod.Commun_11_621
Author(s) : Luo W , Wen Y , Tu Y , Du H , Li Q , Zhu C , Li Y
Ref : Nat Prod Commun , 11 :621 , 2016
Abstract : Ten flavonoids (1-10), including a new glycoside (nevadensin-7-sambubioside, 7), together with a phenylpropanoid glycoside (11) were isolated from Lysionotus pauciflorus. Their structures were elucidated by a combination of spectroscopic methods and comparing with literature data. Five compounds (1, 3, 4, 8, and 9) were obtained from the family Gesneriaceae for the first time. The new compound was evaluated in vitro for anticholinesterase activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), but was found to be inactive.
ESTHER : Luo_2016_Nat.Prod.Commun_11_621
PubMedSearch : Luo_2016_Nat.Prod.Commun_11_621
PubMedID: 27319133

Title : Lysosomal acid lipase regulates VLDL synthesis and insulin sensitivity in mice - Radovic_2016_Diabetologia_59_1743
Author(s) : Radovic B , Vujic N , Leopold C , Schlager S , Goeritzer M , Patankar JV , Korbelius M , Kolb D , Reindl J , Wegscheider M , Tomin T , Birner-Gruenberger R , Schittmayer M , Groschner L , Magnes C , Diwoky C , Frank S , Steyrer E , Du H , Graier WF , Madl T , Kratky D
Ref : Diabetologia , 59 :1743 , 2016
Abstract : AIMS/HYPOTHESIS: Lysosomal acid lipase (LAL) hydrolyses cholesteryl esters and triacylglycerols (TG) within lysosomes to mobilise NEFA and cholesterol. Since LAL-deficient (Lal (-/-) ) mice suffer from progressive loss of adipose tissue and severe accumulation of lipids in hepatic lysosomes, we hypothesised that LAL deficiency triggers alternative energy pathway(s).
METHODS: We studied metabolic adaptations in Lal (-/-) mice.
RESULTS: Despite loss of adipose tissue, Lal (-/-) mice show enhanced glucose clearance during insulin and glucose tolerance tests and have increased uptake of [(3)H]2-deoxy-D-glucose into skeletal muscle compared with wild-type mice. In agreement, fasted Lal (-/-) mice exhibit reduced glucose and glycogen levels in skeletal muscle. We observed 84% decreased plasma leptin levels and significantly reduced hepatic ATP, glucose, glycogen and glutamine concentrations in fed Lal (-/-) mice. Markedly reduced hepatic acyl-CoA concentrations decrease the expression of peroxisome proliferator-activated receptor alpha (PPARalpha) target genes. However, treatment of Lal (-/-) mice with the PPARalpha agonist fenofibrate further decreased plasma TG (and hepatic glucose and glycogen) concentrations in Lal (-/-) mice. Depletion of hepatic nuclear factor 4alpha and forkhead box protein a2 in fasted Lal (-/-) mice might be responsible for reduced expression of microsomal TG transfer protein, defective VLDL synthesis and drastically reduced plasma TG levels. CONCLUSIONS/INTERPRETATION: Our findings indicate that neither activation nor inactivation of PPARalpha per se but rather the availability of hepatic acyl-CoA concentrations regulates VLDL synthesis and subsequent metabolic adaptations in Lal (-/-) mice. We conclude that decreased plasma VLDL production enhances glucose uptake into skeletal muscle to compensate for the lack of energy supply.
ESTHER : Radovic_2016_Diabetologia_59_1743
PubMedSearch : Radovic_2016_Diabetologia_59_1743
PubMedID: 27153842

Title : Lysosomal Acid Lipase Hydrolyzes Retinyl Ester and Affects Retinoid Turnover - Grumet_2016_J.Biol.Chem_291_17977
Author(s) : Grumet L , Eichmann TO , Taschler U , Zierler KA , Leopold C , Moustafa T , Radovic B , Romauch M , Yan C , Du H , Haemmerle G , Zechner R , Fickert P , Kratky D , Zimmermann R , Lass A
Ref : Journal of Biological Chemistry , 291 :17977 , 2016
Abstract : Lysosomal acid lipase (LAL) is essential for the clearance of endocytosed cholesteryl ester and triglyceride-rich chylomicron remnants. Humans and mice with defective or absent LAL activity accumulate large amounts of cholesteryl esters and triglycerides in multiple tissues. Although chylomicrons also contain retinyl esters (REs), a role of LAL in the clearance of endocytosed REs has not been reported. In this study, we found that murine LAL exhibits RE hydrolase activity. Pharmacological inhibition of LAL in the human hepatocyte cell line HepG2, incubated with chylomicrons, led to increased accumulation of REs in endosomal/lysosomal fractions. Furthermore, pharmacological inhibition or genetic ablation of LAL in murine liver largely reduced in vitro acid RE hydrolase activity. Interestingly, LAL-deficient mice exhibited increased RE content in the duodenum and jejunum but decreased RE content in the liver. Furthermore, LAL-deficient mice challenged with RE gavage exhibited largely reduced post-prandial circulating RE content, indicating that LAL is required for efficient nutritional vitamin A availability. In summary, our results indicate that LAL is the major acid RE hydrolase and required for functional retinoid homeostasis.
ESTHER : Grumet_2016_J.Biol.Chem_291_17977
PubMedSearch : Grumet_2016_J.Biol.Chem_291_17977
PubMedID: 27354281
Gene_locus related to this paper: human-LIPA , mouse-1llip

Title : Hepatocyte-Specific Expression of Human Lysosome Acid Lipase Corrects Liver Inflammation and Tumor Metastasis in lal(-\/-) Mice - Du_2015_Am.J.Pathol_185_2379
Author(s) : Du H , Zhao T , Ding X , Yan C
Ref : American Journal of Pathology , 185 :2379 , 2015
Abstract : The liver is a major organ for lipid synthesis and metabolism. Deficiency of lysosomal acid lipase (LAL; official name Lipa, encoded by Lipa) in mice (lal(-/-)) results in enlarged liver size due to neutral lipid storage in hepatocytes and Kupffer cells. To test the functional role of LAL in hepatocyte, hepatocyte-specific expression of human LAL (hLAL) in lal(-/-) mice was established by cross-breeding of liver-activated promoter (LAP)-driven tTA transgene and (tetO)7-CMV-hLAL transgene with lal(-/-) knockout (KO) (LAP-Tg/KO) triple mice. Hepatocyte-specific expression of hLAL in LAP-Tg/KO triple mice reduced the liver size to the normal level by decreasing lipid storage in both hepatocytes and Kupffer cells. hLAL expression reduced tumor-promoting myeloid-derived suppressive cells in the liver of lal(-/-) mice. As a result, B16 melanoma metastasis to the liver was almost completely blocked. Expression and secretion of multiple tumor-promoting cytokines or chemokines in the liver were also significantly reduced. Because hLAL is a secretory protein, lal(-/-) phenotypes in other compartments (eg, blood, spleen, and lung) also ameliorated, including systemic reduction of myeloid-derived suppressive cells, an increase in CD4(+) and CD8(+) T and B lymphocytes, and reduced B16 melanoma metastasis in the lung. These results support a concept that LAL in hepatocytes is a critical metabolic enzyme in controlling neutral lipid metabolism, liver homeostasis, immune response, and tumor metastasis.
ESTHER : Du_2015_Am.J.Pathol_185_2379
PubMedSearch : Du_2015_Am.J.Pathol_185_2379
PubMedID: 26212911
Gene_locus related to this paper: mouse-1llip

Title : Stereoselective inhibition of human butyrylcholinesterase by the enantiomers of bambuterol and their intermediates - Pistolozzi_2015_Drug.Metab.Dispos_43_344
Author(s) : Pistolozzi M , Du H , Wei H , Tan W
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 43 :344 , 2015
Abstract : This work describes the sequential hydrolysis of bambuterol enantiomers and their monocarbamate metabolites (MONO) catalyzed by human butyrylcholinesterase (BChE) as well as the enzyme inhibition resulting from this process. Particular emphasis is given to the contribution given by MONO to the enzyme inhibition because it was not fully characterized in previous works. Bambuterol and MONO enantiomers displayed the same time- and concentration-dependent mechanism of interaction with the enzyme. The hydrolysis kinetics of both bambuterol and MONO was enantioselective, and the (R)-enantiomer of each compound was hydrolyzed fourfold faster than the respective (S)-enantiomer. Even though the enzyme inhibition rates of (R)- and (S)-MONO were much slower than those of their respective bambuterol enantiomers ( approximately 15-fold), both MONO enantiomers showed a significant BChE inhibition when physiologically relevant concentrations of enzyme and inhibitors were used ( approximately 50% of their respective bambuterol enantiomers). The kinetic constants obtained by testing each single compound were used to model the contribution given by MONO to the enzyme inhibition observed for bambuterol. The hydrolysis of MONO enantiomers enhanced the inhibitory power of bambuterol enantiomers of about 27.5% (R) and 12.5% (S) and extended more than 1 hour the duration of inhibition. The data indicate that MONO contribute significantly to the inhibition of BChE occurring in humans upon administration of normal doses of bambuterol. In addition, the hydrolysis of MONO resulted in the rate-limiting step in the conversion of bambuterol in its pharmacologically active metabolite terbutaline; therefore, MONO concentrations should always be monitored during pharmacokinetic studies of bambuterol.
ESTHER : Pistolozzi_2015_Drug.Metab.Dispos_43_344
PubMedSearch : Pistolozzi_2015_Drug.Metab.Dispos_43_344
PubMedID: 25504505

Title : Anticholinesterases and antioxidant alkamides from Piper nigrum fruits - Tu_2015_Nat.Prod.Res__1
Author(s) : Tu Y , Zhong Y , Du H , Luo W , Wen Y , Li Q , Zhu C , Li Y
Ref : Nat Prod Res , :1 , 2015
Abstract : The anticholinesterase and antioxidant effects of five different extracts of Piper nigrum were evaluated. Twenty-one known alkamides were isolated from active ethyl acetate extract and investigated for their cholinesterase inhibitory and antioxidant effects. Among them, piperine (2), piperettine (5) and piperettyline (20) exhibited dual inhibition against AChE and BChE, and feruperine (18) was the most potent selective inhibitor of BChE. Molecular docking simulation was performed to get insight into the binding interactions of the ligands and enzymes. In addition, N-trans-feruloyltyramine (3) contributed to the strongest DPPH radical-scavenging activity. The self-induced Abeta aggregation inhibition of 2, 5 and 18 was further evaluated. Results indicated that some alkamides could be multifunctional lead candidates for Alzheimer's disease therapy.
ESTHER : Tu_2015_Nat.Prod.Res__1
PubMedSearch : Tu_2015_Nat.Prod.Res__1
PubMedID: 26407107

Title : Abundance and significance of neuroligin-1 and glutamate in Hirschsprung's disease - Wang_2015_World.J.Gastroenterol_21_7172
Author(s) : Wang J , Du H , Mou YR , Niu JY , Zhang WT , Yang HC , Li AW
Ref : World J Gastroenterol , 21 :7172 , 2015
Abstract : AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate (Glu) in Hirschsprung's disease (HSCR).
METHODS: Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR (qRT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments (ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay (ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.
RESULTS: Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transitional and aganglionic segments based on Western blot (neuroligin-1: 0.177 +/- 0.008 vs 0.101 +/- 0.006, 0.177 +/- 0.008 vs 0.035 +/- 0.005, and 0.101 +/- 0.006 vs 0.035 +/- 0.005, P < 0.005; Glu: 0.198 +/- 0.006 vs 0.115 +/- 0.008, 0.198 +/- 0.006 vs 0.040 +/- 0.003, and 0.115 +/- 0.008 vs 0.040 +/- 0.003, P < 0.005) and qRT-PCR (neuroligin-1: 9.58 x 10(-5) +/- 9.94 x 10(-6) vs 2.49 x 10(-5) +/- 1.38 x 10(-6), 9.58 x 10(-5) +/- 9.94 x 10(-6) vs 7.17 x 10(-6 +/-) 1.12 x 10(-6), and 2.49 x 10(-5) +/- 1.38 x 10(-6) vs 7.17 x 10(-6) +/- 1.12 x 10(-6), P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA (in nmol/muL, 0.93 +/- 0.31 vs 0.57 +/- 0.25, 0.93 +/- 0.31 vs 0.23 +/- 0.16, and 0.57 +/- 0.25 vs 0.23 +/- 0.16, P < 0.005). CONCLUSION: Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.
ESTHER : Wang_2015_World.J.Gastroenterol_21_7172
PubMedSearch : Wang_2015_World.J.Gastroenterol_21_7172
PubMedID: 26109803

Title : High quality draft genomic sequence of Flavobacterium enshiense DK69(T) and comparison among Flavobacterium genomes - Zeng_2015_Stand.Genomic.Sci_10_92
Author(s) : Zeng Z , Chen C , Du H , Wang G , Li M
Ref : Stand Genomic Sci , 10 :92 , 2015
Abstract : Flavobacterium enshiense DK69(T) is a Gram-negative, aerobic, rod-shaped, non-motile and non-flagellated bacterium that belongs to the family Flavobacteriaceae in the phylum Bacteroidetes. The high quality draft genome of strain DK69(T) was obtained and has a 3,375,260 bp genome size with a G + C content of 37.7 mol % and 2848 protein coding genes. In addition, we sequenced five more genomes of Flavobacterium type strains and performed a comparative genomic analysis among 12 Flavobacterium genomes. The results show some specific genes within the fish pathogenic Flavobacterium strains which provide information for further analysis the pathogenicity.
ESTHER : Zeng_2015_Stand.Genomic.Sci_10_92
PubMedSearch : Zeng_2015_Stand.Genomic.Sci_10_92
PubMedID: 26561515
Gene_locus related to this paper: 9flao-v6scv7

Title : Reversal of advanced disease in lysosomal acid lipase deficient mice: a model for lysosomal acid lipase deficiency disease - Sun_2014_Mol.Genet.Metab_112_229
Author(s) : Sun Y , Xu YH , Du H , Quinn B , Liou B , Stanton L , Inskeep V , Ran H , Jakubowitz P , Grilliot N , Grabowski GA
Ref : Mol Genet Metab , 112 :229 , 2014
Abstract : Lysosomal acid lipase (LAL) is an essential enzyme that hydrolyzes triglycerides (TG) and cholesteryl esters (CE) in lysosomes. Mutations of the LIPA gene lead to Wolman disease (WD) and cholesterol ester storage disease (CESD). The disease hallmarks include hepatosplenomegaly and extensive storage of CE and/or TG. The effects of intravenous investigational enzyme therapy (ET) on survival and efficacy were evaluated in Lipa knock out, lal-/- mice with advanced disease using recombinant human LAL (rhLAL). Comparative ET was conducted with lower doses (weekly, 0.8 and 3.2mg/kg) beginning at 16 weeks (study 1), and with higher dose (10mg/kg) in early (8-weeks), middle (16-weeks) and late (24-weeks) disease stages (study 2). In study 1, rhLAL extended the life span of lal-/- mice in a dose dependent manner by 52 (0.8 mg/kg) or 94 (3.2mg/kg) days. This was accompanied by partial correction of cholesterol and TG levels in spleen and liver. In study 2, the high dose resulted in a significant improvement in organ size (liver, spleen and small intestine) and tissue histology as well as significant decreases in cholesterol and TG in all three groups. In the treated livers and spleens the cholesterol and TG levels were reduced to below treatment initiation levels indicating a reversal of disease manifestations, even in advanced disease. ET diminished liver fibrosis and macrophage proliferation. These results show that LAL deficiency can be improved biochemically and histopathologically by various dosages of ET, even in advanced disease.
ESTHER : Sun_2014_Mol.Genet.Metab_112_229
PubMedSearch : Sun_2014_Mol.Genet.Metab_112_229
PubMedID: 24837159
Gene_locus related to this paper: mouse-1llip

Title : Neuregulin-1 impairs the long-term depression of hippocampal inhibitory synapses by facilitating the degradation of endocannabinoid 2-AG - Du_2013_J.Neurosci_33_15022
Author(s) : Du H , Kwon IK , Kim J
Ref : Journal of Neuroscience , 33 :15022 , 2013
Abstract : Endocannabinoids play essential roles in synaptic plasticity; thus, their dysfunction often causes impairments in memory or cognition. However, it is not well understood whether deficits in the endocannabinoid system account for the cognitive symptoms of schizophrenia. Here, we show that endocannabinoid-mediated synaptic regulation is impaired by the prolonged elevation of neuregulin-1, the abnormality of which is a hallmark in many patients with schizophrenia. When rat hippocampal slices were chronically treated with neuregulin-1, the degradation of 2-arachidonoylglycerol (2-AG), one of the major endocannabinoids, was enhanced due to the increased expression of its degradative enzyme, monoacylglycerol lipase. As a result, the time course of depolarization-induced 2-AG signaling was shortened, and the magnitude of 2-AG-dependent long-term depression of inhibitory synapses was reduced. Our study reveals that an alteration in the signaling of 2-AG contributes to hippocampal synaptic dysfunction in a hyper-neuregulin-1 condition and thus provides novel insights into potential schizophrenic therapeutics that target the endocannabinoid system.
ESTHER : Du_2013_J.Neurosci_33_15022
PubMedSearch : Du_2013_J.Neurosci_33_15022
PubMedID: 24048832

Title : Gene profile of myeloid-derived suppressive cells from the bone marrow of lysosomal acid lipase knock-out mice - Yan_2012_PLoS.One_7_e30701
Author(s) : Yan C , Ding X , Dasgupta N , Wu L , Du H
Ref : PLoS ONE , 7 :e30701 , 2012
Abstract : BACKGROUND: Lysosomal acid lipase (LAL) controls development and homeostasis of myeloid lineage cells. Loss of the lysosomal acid lipase (LAL) function leads to expansion of myeloid-derived suppressive cells (MDSCs) that cause myeloproliferative neoplasm. METHODOLOGY/PRINCIPAL FINDINGS: Affymetrix GeneChip microarray analysis identified detailed intrinsic defects in Ly6G(+) myeloid lineage cells of LAL knock-out (lal-/-) mice. Ingenuity Pathway Analysis revealed activation of the mammalian target of rapamycin (mTOR) signaling, which functions as a nutrient/energy/redox sensor, and controls cell growth, cell cycle entry, cell survival, and cell motility. Loss of the LAL function led to major alteration of large GTPase and small GTPase signal transduction pathways. lal-/- Ly6G(+) myeloid cells in the bone marrow showed substantial increase of cell proliferation in association with up-regulation of cyclin and cyclin-dependent kinase (cdk) genes. The epigenetic microenvironment was significantly changed due to the increased expression of multiple histone cluster genes, centromere protein genes and chromosome modification genes. Gene expression of bioenergetic pathways, including glycolysis, aerobic glycolysis, mitochondrial oxidative phosphorylation, and respiratory chain proteins, was also increased, while the mitochondrial function was impaired in lal-/- Ly6G(+) myeloid cells. The concentration of reactive oxygen species (ROS) was significantly increased accompanied by up-regulation of nitric oxide/ROS production genes in these cells. CONCLUSIONS/SIGNIFICANCE: This comprehensive gene profile study for the first time identifies and defines important gene pathways involved in the myeloid lineage cells towards MDSCs using lal-/- mouse model.
ESTHER : Yan_2012_PLoS.One_7_e30701
PubMedSearch : Yan_2012_PLoS.One_7_e30701
PubMedID: 22383970

Title : Genome sequencing and comparison of two nonhuman primate animal models, the cynomolgus and Chinese rhesus macaques - Yan_2011_Nat.Biotechnol_29_1019
Author(s) : Yan G , Zhang G , Fang X , Zhang Y , Li C , Ling F , Cooper DN , Li Q , Li Y , van Gool AJ , Du H , Chen J , Chen R , Zhang P , Huang Z , Thompson JR , Meng Y , Bai Y , Wang J , Zhuo M , Wang T , Huang Y , Wei L , Li J , Wang Z , Hu H , Yang P , Le L , Stenson PD , Li B , Liu X , Ball EV , An N , Huang Q , Fan W , Zhang X , Wang W , Katze MG , Su B , Nielsen R , Yang H , Wang X
Ref : Nat Biotechnol , 29 :1019 , 2011
Abstract : The nonhuman primates most commonly used in medical research are from the genus Macaca. To better understand the genetic differences between these animal models, we present high-quality draft genome sequences from two macaque species, the cynomolgus/crab-eating macaque and the Chinese rhesus macaque. Comparison with the previously sequenced Indian rhesus macaque reveals that all three macaques maintain abundant genetic heterogeneity, including millions of single-nucleotide substitutions and many insertions, deletions and gross chromosomal rearrangements. By assessing genetic regions with reduced variability, we identify genes in each macaque species that may have experienced positive selection. Genetic divergence patterns suggest that the cynomolgus macaque genome has been shaped by introgression after hybridization with the Chinese rhesus macaque. Macaque genes display a high degree of sequence similarity with human disease gene orthologs and drug targets. However, we identify several putatively dysfunctional genetic differences between the three macaque species, which may explain functional differences between them previously observed in clinical studies.
ESTHER : Yan_2011_Nat.Biotechnol_29_1019
PubMedSearch : Yan_2011_Nat.Biotechnol_29_1019
PubMedID: 22002653
Gene_locus related to this paper: macfa-BCHE , macfa-g7nzc0 , macfa-g7nze2 , macfa-g7p4b9 , macfa-g7pa87 , macfa-g7pd01 , macfa-g7q259 , macfa-3neur , macfa-g8f585 , macfa-KANSL3 , macfa-q4r8p0 , macfa-SPG21 , macfa-TEX30 , macmu-3neur , macmu-ACHE , macmu-BCHE , macmu-f6sz31 , macmu-f6the6 , macmu-f6zkq5 , macmu-f7buk8 , macmu-f7cfi8 , macmu-f7flv1 , macmu-f7ggk1 , macmu-f7hir7 , macmu-g7n054 , macmu-g7npb8 , macmu-g7nq39 , macmu-KANSL3 , macmu-TEX30 , macfa-g7pgg6 , macmu-g7n4x3 , macfa-g7nzx2 , macfa-g8f4f7 , macmu-f7ba84 , macfa-g7psx7 , macmu-h9er02 , macfa-g8f3k0 , macfa-a0a2k5w1n7 , macmu-g7mxj6 , macfa-g7pbk1 , macfa-a0a2k5urk5 , macfa-a0a2k5wye4 , macfa-g7pe14 , macmu-f7hkw9 , macmu-f7hm08 , macmu-g7mke4 , macfa-g7nxn9 , macmu-a0a1d5rh04 , macmu-h9fud6 , macfa-g8f3e1 , macfa-i7gcw6 , macmu-f6qwx1 , macmu-f7h4t2 , macfa-a0a2k5wkd0 , macfa-a0a2k5v7v4 , macfa-g7p7y3 , macfa-a0a2k5uqq3 , macmu-i2cu80 , macfa-g8f5i1 , macmu-f7h550 , macmu-f7gkb9 , macfa-a0a2k5tui1

Title : Lysosomal acid lipase deficiency impairs regulation of ABCA1 gene and formation of high density lipoproteins in cholesteryl ester storage disease - Bowden_2011_J.Biol.Chem_286_30624
Author(s) : Bowden KL , Bilbey NJ , Bilawchuk LM , Boadu E , Sidhu R , Ory DS , Du H , Chan T , Francis GA
Ref : Journal of Biological Chemistry , 286 :30624 , 2011
Abstract : ATP-binding cassette transporter A1 (ABCA1) mediates the rate-limiting step in high density lipoprotein (HDL) particle formation, and its expression is regulated primarily by oxysterol-dependent activation of liver X receptors. We previously reported that ABCA1 expression and HDL formation are impaired in the lysosomal cholesterol storage disorder Niemann-Pick disease type C1 and that plasma HDL-C is low in the majority of Niemann-Pick disease type C patients. Here, we show that ABCA1 regulation and activity are also impaired in cholesteryl ester storage disease (CESD), caused by mutations in the LIPA gene that result in less than 5% of normal lysosomal acid lipase (LAL) activity. Fibroblasts from patients with CESD showed impaired up-regulation of ABCA1 in response to low density lipoprotein (LDL) loading, reduced phospholipid and cholesterol efflux to apolipoprotein A-I, and reduced alpha-HDL particle formation. Treatment of normal fibroblasts with chloroquine to inhibit LAL activity reduced ABCA1 expression and activity, similar to that of CESD cells. Liver X receptor agonist treatment of CESD cells corrected ABCA1 expression but failed to correct LDL cholesteryl ester hydrolysis and cholesterol efflux to apoA-I. LDL-induced production of 27-hydroxycholesterol was reduced in CESD compared with normal fibroblasts. Treatment with conditioned medium containing LAL from normal fibroblasts or with recombinant human LAL rescued ABCA1 expression, apoA-I-mediated cholesterol efflux, HDL particle formation, and production of 27-hydroxycholesterol by CESD cells. These results provide further evidence that the rate of release of cholesterol from late endosomes/lysosomes is a critical regulator of ABCA1 expression and activity, and an explanation for the hypoalphalipoproteinemia seen in CESD patients.
ESTHER : Bowden_2011_J.Biol.Chem_286_30624
PubMedSearch : Bowden_2011_J.Biol.Chem_286_30624
PubMedID: 21757691
Gene_locus related to this paper: human-LIPA

Title : Critical roles of lysosomal acid lipase in myelopoiesis - Qu_2010_Am.J.Pathol_176_2394
Author(s) : Qu P , Shelley WC , Yoder MC , Wu L , Du H , Yan C
Ref : American Journal of Pathology , 176 :2394 , 2010
Abstract : Lysosomal acid lipase (LAL) is a key enzyme that cleaves cholesteryl esters and triglycerides to generate free fatty acids and cholesterol in lysosomes. Genetic ablation of the lal gene (lal(-/-)) in mice has resulted in a systemic increase of macrophages and neutrophils, causing severe inflammation and pathogenesis in multiple organs. We hypothesized that aberrant growth and differentiation of myeloid cells in lal(-/-) mice arises from dysregulated production of progenitor cells in the bone marrow. Indeed, lal(-/-) mice displayed increased numbers of primitive lin(-)Sca-1(+)c-Kit(+) (LSK) cells and granulocyte-macrophage precursors (GMP). Increased high proliferative potential colony-forming cells (HPP-CFC) were enumerated from cultured lal(-/-) bone marrow cells, as were significantly more CFU-GM, CFU-G, and CFU-M colonies. As a consequence, lal(-/-) mice developed significant myeloid infiltration, particularly with CD11b+/Gr-1+ myeloid-derived suppressive cells in multiple organs. Both decreased apoptosis and increased proliferation contribute to the systemic increase of myeloid cells in lal(-/-) myeloid cells. These lal(-/-) CD11b(+)/Gr-1(+) cells displayed suppressive activity on T cell proliferation and function in vitro. Bone marrow chimeras confirmed that the myeloproliferative disorder in lal(-/-) mice was primarily attributable to autonomous defects in myeloid progenitor cells, although the hematopoietic microenvironment in the lal(-/-) mice did not support hematopoiesis normally. These results provide evidence that LAL is an important regulator of myelopoiesis during hematopoietic development, differentiation, and homeostasis.
ESTHER : Qu_2010_Am.J.Pathol_176_2394
PubMedSearch : Qu_2010_Am.J.Pathol_176_2394
PubMedID: 20348241
Gene_locus related to this paper: mouse-1llip

Title : Critical roles of lysosomal acid lipase in T cell development and function - Qu_2009_Am.J.Pathol_174_944
Author(s) : Qu P , Du H , Wilkes DS , Yan C
Ref : American Journal of Pathology , 174 :944 , 2009
Abstract : Lysosomal acid lipase (LAL) cleaves cholesteryl esters and triglycerides to generate free fatty acids and cholesterol in lysosomes. In LAL gene-knockout (lal(-/-)) mice, blockage of cholesteryl ester and triglyceride metabolism led to abnormal organization of the thymus and spleen, as well as neutral lipid accumulation in these organs. LAL deficiency impaired T cell development in the thymus. Peripheral T cells were reduced dramatically in lal(-/-) mice, due largely to increased apoptosis and decreased proliferation of lal(-/-) T cells in the thymus and peripheral compartments. These lal(-/-) T cells lost the ability to respond to T cell receptor stimulation, including reduced expression of cell surface receptor CD69, abolishment of T cell proliferation, and decreased expression of T lymphokines after stimulation by either anti-CD3 plus anti-CD28 or phorbol-12-myristate-13-acetate and ionomycin. Differentiation of Th1 and Th2 CD4(+) effector lymphocytes by T cell receptor stimulation was blocked in lal(-/-) mice. The ratio of CD4(+)CD25(+)FoxP3(+) Tregs to CD4(+) T cells was increased in lal(-/-) spleens. Bone marrow chimeras demonstrated retardation of T cell development and maturation in lal(-/-) mice due to defects in T cell precursors. Therefore, LAL, its downstream genes, and lipid mediators all play essential roles in development, homeostasis, and function of T cells. The altered development and function of lal(-/-) T cells contributes to disease formation in various organs during LAL deficiency.
ESTHER : Qu_2009_Am.J.Pathol_174_944
PubMedSearch : Qu_2009_Am.J.Pathol_174_944
PubMedID: 19179613
Gene_locus related to this paper: mouse-1llip

Title : Clinical, experimental, and genomic differences between intermediately pathogenic, highly pathogenic, and epidemic Streptococcus suis - Ye_2009_J.Infect.Dis_199_97
Author(s) : Ye C , Zheng H , Zhang J , Jing H , Wang L , Xiong Y , Wang W , Zhou Z , Sun Q , Luo X , Du H , Gottschalk M , Xu J
Ref : J Infect Dis , 199 :97 , 2009
Abstract : BACKGROUND: Streptococcus suis emerged to cause an unusual outbreak of streptococcal toxic-shock-like syndrome (STSLS) in 2005. The mechanisms involved are unknown. METHODS: Clinical, laboratory, and epidemiologic data on patients infected with culture-confirmed S. suis were analyzed. The strain involved in the outbreak, "epidemic" strain ST7, was compared with both a classical highly pathogenic strain, ST1, and an intermediately pathogenic strain, ST25, to determine both its capacity to induce cytokines in experimentally infected mice and its genomic difference. RESULTS: Of 38 patients infected with culture-confirmed S. suis, 14 presented with STSLS. During the early phase of the disease, serum levels of interleukin (IL)-1beta, IL-6, IL-8, IL-12p70, interferon-gamma, and tumor necrosis factor-alpha were more elevated in patients with STSLS than in those with meningitis only. Serum levels of proinflammatory cytokines were significantly higher in mice infected with ST7 than in those infected with either ST1 or ST25. Genomic comparisons with ST25 showed that ST1 had acquired 132 genomic islands, including 5 pathogenicity islands, and that ST7, the epidemic strain, had acquired an additional 5 genomic islands. CONCLUSION: Intermediately pathogenic strain ST25 has evolved to become highly pathogenic strain ST1, which, in turn, has more recently evolved to become epidemic strain ST7. ST7 has the ability to stimulate the production of massive amounts of proinflammatory cytokines, leading to STSLS.
ESTHER : Ye_2009_J.Infect.Dis_199_97
PubMedSearch : Ye_2009_J.Infect.Dis_199_97
PubMedID: 19016627
Gene_locus related to this paper: strsu-a4vws4 , strsu-q673u2 , strsy-a4vus4

Title : Chemical screen to reduce sterol accumulation in Niemann-Pick C disease cells identifies novel lysosomal acid lipase inhibitors - Rosenbaum_2009_Biochim.Biophys.Acta_1791_1155
Author(s) : Rosenbaum AI , Rujoi M , Huang AY , Du H , Grabowski GA , Maxfield FR
Ref : Biochimica & Biophysica Acta , 1791 :1155 , 2009
Abstract : Niemann-Pick C disease (NPC) is a lysosomal storage disorder causing abnormal accumulation of unesterified free cholesterol in lysosomal storage organelles. High content phenotypic microscopy chemical screens in both human and hamster NPC-deficient cells have identified several compounds that partially revert the NPC phenotype. Cell biological and biochemical studies show that several of these molecules inhibit lysosomal acid lipase, the enzyme that hydrolyzes LDL-derived triacylglycerol and cholesteryl esters. The effects of reduced lysosomal acid lipase activity in lowering cholesterol accumulation in NPC mutant cells were verified by RNAi-mediated knockdown of lysosomal acid lipase in NPC1-deficient human fibroblasts. This work demonstrates the utility of phenotypic cellular screens as a means to identify molecular targets for altering a complex process such as intracellular cholesterol trafficking and metabolism.
ESTHER : Rosenbaum_2009_Biochim.Biophys.Acta_1791_1155
PubMedSearch : Rosenbaum_2009_Biochim.Biophys.Acta_1791_1155
PubMedID: 19699313

Title : Effects of 3-benzidino-6-phenylpyridazine, as an acetylcholinesterase inhibitor, on outward potassium current in acutely isolated rat hippocampal pyramidal neurons - Du_2008_Toxicol.Lett_181_104
Author(s) : Du H , Li M , Yang P
Ref : Toxicol Lett , 181 :104 , 2008
Abstract : As an acetylcholinesterase (AChE) inhibitor, the effects of 3-benzidino-6-phenylpyridazine (BPP) on outward potassium current including delayed rectifier potassium current (I(K(DR))) and transient outward potassium current (I(K(A))) in acutely isolated rat hippocampal pyramidal neurons were studied, using the whole cell patch-clamp technique. BPP reversibly inhibited electric eel AChE as an inhibitor, with IC(50) of 1.43 microM. BPP (0.10-100 microM) decreased I(K(DR)) and I(K(A)) in a concentration-dependent, voltage-independent and partial reversible manner, with IC(50) of 0.47 and 0.31 microM, respectively. 10 microM BPP did not affect steady-state activation of I(K(DR)) and I(K(A)). In addition, 10 microM BPP shifted the voltage dependence of steady-state inactivation of I(K(A)) towards negative potential. In conclusion, BPP potently inhibits I(K(DR)) and I(K(A)) in rat hippocampal pyramidal neurons, which may contribute to BPP's restoring the damaged central nervous system.
ESTHER : Du_2008_Toxicol.Lett_181_104
PubMedSearch : Du_2008_Toxicol.Lett_181_104
PubMedID: 18675331

Title : Wolman disease\/cholesteryl ester storage disease: efficacy of plant-produced human lysosomal acid lipase in mice - Du_2008_J.Lipid.Res_49_1646
Author(s) : Du H , Cameron TL , Garger SJ , Pogue GP , Hamm LA , White E , Hanley KM , Grabowski GA
Ref : J Lipid Res , 49 :1646 , 2008
Abstract : Lysosomal acid lipase (LAL) is an essential enzyme that hydrolyzes triglycerides (TGs) and cholesteryl esters (CEs) in lysosomes. Genetic LAL mutations lead to Wolman disease (WD) and cholesteryl ester storage disease (CESD). An LAL-null (lal(-/-)) mouse model resembles human WD/CESD with storage of CEs and TGs in multiple organs. Human LAL (hLAL) was expressed in Nicotiana benthamiana using the GENEWARE expression system (G-hLAL). Purified G-hLAL showed mannose receptor-dependent uptake into macrophage cell lines (J774E). Intraperitoneal injection of G-hLAL produced peak activities in plasma at 60 min and in the liver and spleen at 240 min. The t(1/2) values were: approximately 90 min (plasma), approximately 14 h (liver), and approximately 32 h (spleen), with return to baseline by approximately 150 h in liver and approximately 200 h in spleen. Ten injections of G-hLAL (every 3 days) into lal(-/-) mice produced normalization of hepatic color, decreases in hepatic cholesterol and TG contents, and diminished foamy macrophages in liver, spleen, and intestinal villi. All injected lal(-/-) mice developed anti-hLAL protein antibodies, but suffered no adverse events. These studies demonstrate the feasibility of using plant-expressed, recombinant hLAL for the enzyme therapy of human WD/CESD with general implications for other lysosomal storage diseases.
ESTHER : Du_2008_J.Lipid.Res_49_1646
PubMedSearch : Du_2008_J.Lipid.Res_49_1646
PubMedID: 18413899
Gene_locus related to this paper: mouse-1llip

Title : Lysosomal acid lipase over-expression disrupts lamellar body genesis and alveolar structure in the lung - Li_2007_Int.J.Exp.Pathol_88_427
Author(s) : Li Y , Qin Y , Li H , Wu R , Yan C , Du H
Ref : International Journal of Experimental Pathology , 88 :427 , 2007
Abstract : The functional role of neutral lipids in the lung is poorly understood. Lysosomal acid lipase (LAL) is a critical enzyme in hydrolysis of cholesteryl esters and triglycerides to generate free fatty acids and cholesterol in lysosomes. Human LAL was over-expressed in a doxycycline-controlled system in mouse respiratory epithelial cells to accelerate intracellular neutral lipid degradation and perturb the surfactant homeostasis in the lung. In this animal system, neutral lipid concentrations of pulmonary surfactant were reduced in bronchoalveolar lavage fluid (BALF) in association with decrease of surfactant protein C (SP-C) gene expression. The size and the number of lamellar bodies in alveolar type II epithelial cells (AT II cells) were significantly reduced accordingly. The number of macrophages required for surfactant recycling in BALF was also significantly reduced. As a result of these combinatory effects, emphysema of the alveolar structure was observed. Taken together, neutral lipid homeostasis is essential for maintenance of lamellar body genesis and the alveolar structure in the lung.
ESTHER : Li_2007_Int.J.Exp.Pathol_88_427
PubMedSearch : Li_2007_Int.J.Exp.Pathol_88_427
PubMedID: 18039279
Gene_locus related to this paper: mouse-1llip

Title : Gene delivery by the hSP-B promoter to lung alveolar type II epithelial cells in LAL-knockout mice through bone marrow mesenchymal stem cells - Yan_2007_Gene.Ther_14_1461
Author(s) : Yan C , Lian X , Dai Y , Wang X , Qu P , White A , Qin Y , Du H
Ref : Gene Therapy , 14 :1461 , 2007
Abstract : Tissue damage and inflammation promote bone marrow stem cells (BMSCs) to differentiate into a variety of cell types in residing tissues. BMSCs can stably maintain their plasticity and are an ideal cell population for delivery of therapeutic genes to non-hematopoietic tissues. Using lacZ as a reporter gene, we demonstrated that the lung-specific human surfactant protein B (hSP-B) 1.5-kb promoter is able to deliver the lacZ gene into the lung of lysosomal acid lipase (LAL) gene-knockout (lal-/-) mice by beta-galactosidase staining, flow cytometry and double immunofluorescence staining. Around 10-18% alveolar type II epithelial cells (AT II cells) exhibited positive lacZ gene expression after 8 weeks of BMSC injection in recipient lal-/- mice. The wild-type mice exhibited no expression after the same treatment. BMSCs from hSP-B 1.5-kb lacZ transgenic mice entered and repopulated in lal-/- bone marrow. The study supports a concept that pulmonary inflammation caused by LAL deficiency can trigger BMSC residing in lal-/- bone marrow, migrating into the lung and converting into residential AT II cells. The hSP-B 1.5 kb promoter is an ideal tool to deliver therapeutic genes into AT II cells through BMSCs to cure pulmonary inflammation-triggered diseases.
ESTHER : Yan_2007_Gene.Ther_14_1461
PubMedSearch : Yan_2007_Gene.Ther_14_1461
PubMedID: 17700706

Title : Macrophage-specific expression of human lysosomal acid lipase corrects inflammation and pathogenic phenotypes in lal-\/- mice - Yan_2006_Am.J.Pathol_169_916
Author(s) : Yan C , Lian X , Li Y , Dai Y , White A , Qin Y , Li H , Hume DA , Du H
Ref : American Journal of Pathology , 169 :916 , 2006
Abstract : Lysosomal acid lipase (LAL) hydrolyzes cholesteryl esters and triglycerides to generate free fatty acids and cholesterol in the cell. The downstream metabolites of these compounds serve as hormonal ligands for nuclear receptors and transcription factors. Genetic ablation of the lal gene in the mouse caused malformation of macrophages and inflammation-triggered multiple pathogenic phenotypes in multiple organs. To assess the relationship between macrophages and lal-/- pathogenic phenotypes, a macrophage-specific doxycycline-inducible transgenic system was generated to induce human LAL (hLAL) expression in the lal-/- genetic background under control of the 7.2-kb c-fms promoter/intron2 regulatory sequence. Doxycycline-induced hLAL expression in macrophages significantly ameliorated aberrant gene expression, inflammatory cell (neutrophil) influx, and pathogenesis in multiple organs. These studies strongly support that neutral lipid metabolism in macrophages contributes to organ inflammation and pathogenesis.
ESTHER : Yan_2006_Am.J.Pathol_169_916
PubMedSearch : Yan_2006_Am.J.Pathol_169_916
PubMedID: 16936266

Title : Neutral lipids and peroxisome proliferator-activated receptor-{gamma} control pulmonary gene expression and inflammation-triggered pathogenesis in lysosomal acid lipase knockout mice - Lian_2005_Am.J.Pathol_167_813
Author(s) : Lian X , Yan C , Qin Y , Knox L , Li T , Du H
Ref : American Journal of Pathology , 167 :813 , 2005
Abstract : The functional roles of neutral lipids in the lung are poorly understood. However, blocking cholesteryl ester and triglyceride metabolism in lysosomal acid lipase gene knockout mice (lal-/-) results in severe pathogenic phenotypes in the lung, including massive neutrophil infiltration, foamy macrophage accumulation, unwanted cell growth, and emphysema. To elucidate the mechanism underlining these pathologies, we performed Affymetrix GeneChip microarray analysis of 1-, 3-, and 6-month-old mice and identified aberrant gene expression that progressed with age. Among changed genes, matrix metalloproteinase (MMP)-12, apoptosis inhibitor 6 (Api-6), erythroblast transformation-specific domain (Ets) transcription factor family member Spi-C, and oncogene MafB were increased 100-, 70-, 40-, and 10-fold, respectively, in lal-/- lungs versus the wild-type lungs. The pathogenic increases of these molecules occurred primarily in alveolar type II epithelial cells. Transcriptional activities of the MMP-12 and Api-6 promoters were stimulated by Spi-C or MafB in respiratory epithelial cells. Treatment with 9-hydroxyoctadecanoic acids and ciglitazone significantly rescued lal-/- pulmonary inflammation and aberrant gene expression. In addition, both compounds as well as peroxisome proliferator-activated receptor gamma inhibited MMP-12 and Api-6 promoter activities. These data suggest that inflammation-triggered cell growth and emphysema during lysosomal acid lipase deficiency are partially caused by peroxisome proliferator-activated receptor-gamma inactivation.
ESTHER : Lian_2005_Am.J.Pathol_167_813
PubMedSearch : Lian_2005_Am.J.Pathol_167_813
PubMedID: 16127159
Gene_locus related to this paper: mouse-1llip

Title : Generation and annotation of the DNA sequences of human chromosomes 2 and 4 - Hillier_2005_Nature_434_724
Author(s) : Hillier LW , Graves TA , Fulton RS , Fulton LA , Pepin KH , Minx P , Wagner-McPherson C , Layman D , Wylie K , Sekhon M , Becker MC , Fewell GA , Delehaunty KD , Miner TL , Nash WE , Kremitzki C , Oddy L , Du H , Sun H , Bradshaw-Cordum H , Ali J , Carter J , Cordes M , Harris A , Isak A , Van Brunt A , Nguyen C , Du F , Courtney L , Kalicki J , Ozersky P , Abbott S , Armstrong J , Belter EA , Caruso L , Cedroni M , Cotton M , Davidson T , Desai A , Elliott G , Erb T , Fronick C , Gaige T , Haakenson W , Haglund K , Holmes A , Harkins R , Kim K , Kruchowski SS , Strong CM , Grewal N , Goyea E , Hou S , Levy A , Martinka S , Mead K , McLellan MD , Meyer R , Randall-Maher J , Tomlinson C , Dauphin-Kohlberg S , Kozlowicz-Reilly A , Shah N , Swearengen-Shahid S , Snider J , Strong JT , Thompson J , Yoakum M , Leonard S , Pearman C , Trani L , Radionenko M , Waligorski JE , Wang C , Rock SM , Tin-Wollam AM , Maupin R , Latreille P , Wendl MC , Yang SP , Pohl C , Wallis JW , Spieth J , Bieri TA , Berkowicz N , Nelson JO , Osborne J , Ding L , Sabo A , Shotland Y , Sinha P , Wohldmann PE , Cook LL , Hickenbotham MT , Eldred J , Williams D , Jones TA , She X , Ciccarelli FD , Izaurralde E , Taylor J , Schmutz J , Myers RM , Cox DR , Huang X , McPherson JD , Mardis ER , Clifton SW , Warren WC , Chinwalla AT , Eddy SR , Marra MA , Ovcharenko I , Furey TS , Miller W , Eichler EE , Bork P , Suyama M , Torrents D , Waterston RH , Wilson RK
Ref : Nature , 434 :724 , 2005
Abstract : Human chromosome 2 is unique to the human lineage in being the product of a head-to-head fusion of two intermediate-sized ancestral chromosomes. Chromosome 4 has received attention primarily related to the search for the Huntington's disease gene, but also for genes associated with Wolf-Hirschhorn syndrome, polycystic kidney disease and a form of muscular dystrophy. Here we present approximately 237 million base pairs of sequence for chromosome 2, and 186 million base pairs for chromosome 4, representing more than 99.6% of their euchromatic sequences. Our initial analyses have identified 1,346 protein-coding genes and 1,239 pseudogenes on chromosome 2, and 796 protein-coding genes and 778 pseudogenes on chromosome 4. Extensive analyses confirm the underlying construction of the sequence, and expand our understanding of the structure and evolution of mammalian chromosomes, including gene deserts, segmental duplications and highly variant regions.
ESTHER : Hillier_2005_Nature_434_724
PubMedSearch : Hillier_2005_Nature_434_724
PubMedID: 15815621
Gene_locus related to this paper: human-ABHD1 , human-LDAH , human-ABHD18 , human-KANSL3 , human-PGAP1 , human-PREPL

Title : MRI of fat distribution in a mouse model of lysosomal acid lipase deficiency - Du_2005_AJR.Am.J.Roentgenol_184_658
Author(s) : Du H , Dardzinski BJ , O'Brien KJ , Donnelly LF
Ref : AJR Am J Roentgenol , 184 :658 , 2005
Abstract : OBJECTIVE: We assessed the use of MRI in the evaluation of abdominal fat distribution in a lysosomal acid lipase (LAL)-deficient mouse model. MATERIALS AND
METHODS: LAL-deficient mice are born with a normal fat distribution but over time deplete the fat stores in the subcutaneous and retroperitoneal tissues and accumulate fat in the liver, spleen, and bowel. Four MRI studies of LAL-deficient mice and control mice were obtained with 3-T T1-weighted spin-echo images and volume segmentation processing to create parameters for the study of fat distribution: intraabdominal adipose tissue-subcutaneous adipose tissue (IAT/SAT) ratio, liver volume, reproductive fat, and retroperitoneal fat. MRI adiposity parameters in LAL-deficient mice were compared with those in control mice. Adiposity volumes calculated on MRI were compared with those calculated at autopsy.
RESULTS: Statistically significant differences were found between LAL-deficient and control mice for IAT/SAT ratio (p=0.0336), liver volume (p=0.0336), and reproductive fat (p=0.0336), and a statistically significant trend was found for retroperitoneal fat (p=0.0514). No statistically significant difference was found between adiposity volumes calculated on MRI and adiposity volumes found at autopsy (all p >0.2). CONCLUSION: Use of an in vivo model showed MRI techniques to be accurate in predicting visceral adiposity. LAL-deficient mice provided a unique model showing a pattern of adipose distribution that is markedly different from that in control mice, and MRI may provide a means of evaluating therapeutic interventions sequentially.
ESTHER : Du_2005_AJR.Am.J.Roentgenol_184_658
PubMedSearch : Du_2005_AJR.Am.J.Roentgenol_184_658
PubMedID: 15671394

Title : Lysosomal acid lipase deficiency causes respiratory inflammation and destruction in the lung - Lian_2004_Am.J.Physiol.Lung.Cell.Mol.Physiol_286_L801
Author(s) : Lian X , Yan C , Yang L , Xu Y , Du H
Ref : American Journal of Physiology Lung Cell Mol Physiol , 286 :L801 , 2004
Abstract : The functional roles of neutral lipids are poorly understood in the lung. Blocking cholesteryl ester and triglyceride metabolism in lysosomal acid lipase gene knockout mice (lal-/-) resulted in a high level of neutrophil influx in the lungs as early as 2 mo of age. Bronchoalveolar macrophages appeared foamy and gradually increased in number with age progression. Affymetrix GeneChip array analysis of lung mRNA showed increased levels of proinflammatory cytokine (including IL-1beta, IL-6, and TNF-alpha) and matrix metalloproteinase (including MMP-8, MMP-9, and MMP-12) expression in lal-/- mice. With age progression, some areas of lal-/- mice developed severe abnormal cell proliferation and alveolar remodeling. In other areas, alveolar destruction (i.e., emphysema) was observed. In addition, Clara cell hypertrophy and hyperplasia developed in conducting airways. The pathophysiological phenotypes in the lal-/- mouse lungs became more severe with increasing age. The studies support the concept that neutral lipid metabolites play essential roles in pulmonary homeostasis, inflammatory responses, remodeling, and injury repair.
ESTHER : Lian_2004_Am.J.Physiol.Lung.Cell.Mol.Physiol_286_L801
PubMedSearch : Lian_2004_Am.J.Physiol.Lung.Cell.Mol.Physiol_286_L801
PubMedID: 14644759
Gene_locus related to this paper: mouse-1llip

Title : The DNA sequence of human chromosome 7 - Hillier_2003_Nature_424_157
Author(s) : Hillier LW , Fulton RS , Fulton LA , Graves TA , Pepin KH , Wagner-McPherson C , Layman D , Maas J , Jaeger S , Walker R , Wylie K , Sekhon M , Becker MC , O'Laughlin MD , Schaller ME , Fewell GA , Delehaunty KD , Miner TL , Nash WE , Cordes M , Du H , Sun H , Edwards J , Bradshaw-Cordum H , Ali J , Andrews S , Isak A , Vanbrunt A , Nguyen C , Du F , Lamar B , Courtney L , Kalicki J , Ozersky P , Bielicki L , Scott K , Holmes A , Harkins R , Harris A , Strong CM , Hou S , Tomlinson C , Dauphin-Kohlberg S , Kozlowicz-Reilly A , Leonard S , Rohlfing T , Rock SM , Tin-Wollam AM , Abbott A , Minx P , Maupin R , Strowmatt C , Latreille P , Miller N , Johnson D , Murray J , Woessner JP , Wendl MC , Yang SP , Schultz BR , Wallis JW , Spieth J , Bieri TA , Nelson JO , Berkowicz N , Wohldmann PE , Cook LL , Hickenbotham MT , Eldred J , Williams D , Bedell JA , Mardis ER , Clifton SW , Chissoe SL , Marra MA , Raymond C , Haugen E , Gillett W , Zhou Y , James R , Phelps K , Iadanoto S , Bubb K , Simms E , Levy R , Clendenning J , Kaul R , Kent WJ , Furey TS , Baertsch RA , Brent MR , Keibler E , Flicek P , Bork P , Suyama M , Bailey JA , Portnoy ME , Torrents D , Chinwalla AT , Gish WR , Eddy SR , McPherson JD , Olson MV , Eichler EE , Green ED , Waterston RH , Wilson RK
Ref : Nature , 424 :157 , 2003
Abstract : Human chromosome 7 has historically received prominent attention in the human genetics community, primarily related to the search for the cystic fibrosis gene and the frequent cytogenetic changes associated with various forms of cancer. Here we present more than 153 million base pairs representing 99.4% of the euchromatic sequence of chromosome 7, the first metacentric chromosome completed so far. The sequence has excellent concordance with previously established physical and genetic maps, and it exhibits an unusual amount of segmentally duplicated sequence (8.2%), with marked differences between the two arms. Our initial analyses have identified 1,150 protein-coding genes, 605 of which have been confirmed by complementary DNA sequences, and an additional 941 pseudogenes. Of genes confirmed by transcript sequences, some are polymorphic for mutations that disrupt the reading frame.
ESTHER : Hillier_2003_Nature_424_157
PubMedSearch : Hillier_2003_Nature_424_157
PubMedID: 12853948
Gene_locus related to this paper: human-ABHD11 , human-ACHE , human-CPVL , human-DPP6 , human-MEST

Title : The male-specific region of the human Y chromosome is a mosaic of discrete sequence classes - Skaletsky_2003_Nature_423_825
Author(s) : Skaletsky H , Kuroda-Kawaguchi T , Minx PJ , Cordum HS , Hillier L , Brown LG , Repping S , Pyntikova T , Ali J , Bieri T , Chinwalla A , Delehaunty A , Delehaunty K , Du H , Fewell G , Fulton L , Fulton R , Graves T , Hou SF , Latrielle P , Leonard S , Mardis E , Maupin R , McPherson J , Miner T , Nash W , Nguyen C , Ozersky P , Pepin K , Rock S , Rohlfing T , Scott K , Schultz B , Strong C , Tin-Wollam A , Yang SP , Waterston RH , Wilson RK , Rozen S , Page DC
Ref : Nature , 423 :825 , 2003
Abstract : The male-specific region of the Y chromosome, the MSY, differentiates the sexes and comprises 95% of the chromosome's length. Here, we report that the MSY is a mosaic of heterochromatic sequences and three classes of euchromatic sequences: X-transposed, X-degenerate and ampliconic. These classes contain all 156 known transcription units, which include 78 protein-coding genes that collectively encode 27 distinct proteins. The X-transposed sequences exhibit 99% identity to the X chromosome. The X-degenerate sequences are remnants of ancient autosomes from which the modern X and Y chromosomes evolved. The ampliconic class includes large regions (about 30% of the MSY euchromatin) where sequence pairs show greater than 99.9% identity, which is maintained by frequent gene conversion (non-reciprocal transfer). The most prominent features here are eight massive palindromes, at least six of which contain testis genes.
ESTHER : Skaletsky_2003_Nature_423_825
PubMedSearch : Skaletsky_2003_Nature_423_825
PubMedID: 12815422
Gene_locus related to this paper: human-NLGN4Y

Title : Lysosomal Acid lipase deficiency: correction of lipid storage by adenovirus-mediated gene transfer in mice - Du_2002_Hum.Gene.Ther_13_1361
Author(s) : Du H , Heur M , Witte DP , Ameis D , Grabowski GA
Ref : Hum Gene Therapy , 13 :1361 , 2002
Abstract : Lysosomal acid lipase (LAL) is the essential enzyme for hydrolysis of triglycerides (TGs) and cholesteryl esters (CEs) in lysosomes. Its deficiency produces two human phenotypes: Wolman disease (WD) and cholesteryl ester storage disease (CESD). The LAL null (lal(-/-)) mouse mimicks aspects of human WD and CESD. The potential for gene therapy of LAL deficiency was tested with first-generation adenoviral vectors containing human LAL cDNA (Ad-hLAL) by intravenous injection into lal(-/-) mice. Compared with phosphate-buffered saline-injected controls, the mice receiving Ad-hLAL had increased hepatic LAL activity, decreased hepatomegaly, and normalization of histopathology. hLAL protein and mRNA were detected by immunohistochemical staining and in situ hybridization in hepatic parenchymal and sinusoid lining cells, splenic sinusoidal cells, lung macrophages, and adrenal cortical cells. Mice showed TG reductions in liver, spleen, and small intestine of 68, 54, and 50%, respectively, and cholesterol reductions of 55, 52, and 34%, respectively, at 20 days postinjection. These studies provide the basis for the use of gene therapy, in the form of gene transfer via intravenously administered adenovirus, to correct deficiency states, such as WD and CESD, and histopathology of a variety of tissues.
ESTHER : Du_2002_Hum.Gene.Ther_13_1361
PubMedSearch : Du_2002_Hum.Gene.Ther_13_1361
PubMedID: 12162818
Gene_locus related to this paper: mouse-1llip

Title : Phenotypic correction of lipid storage and growth arrest in wolman disease fibroblasts by gene transfer of lysosomal acid lipase - Tietge_2001_Hum.Gene.Ther_12_279
Author(s) : Tietge UJ , Sun G , Czarnecki S , Yu Q , Lohse P , Du H , Grabowski GA , Glick JM , Rader DJ
Ref : Hum Gene Therapy , 12 :279 , 2001
Abstract : Wolman disease is a lethal lysosomal storage disease due to deficiency of lysosomal acid lipase (LAL). Wolman disease is characterized by pronounced hepatic involvement while neurological symptoms are uncommon, making Wolman disease an attractive candidate for liver-directed gene therapy. This study was performed to test the effects of gene replacement in fibroblasts lacking LAL, using a recombinant adenovirus encoding the human LAL cDNA (AdhLAL). Human fibroblasts from a Wolman disease patient were infected with AdhLAL and showed a dose-dependent increase in LAL protein and activity up to 5-fold above levels in control fibroblasts. Furthermore, 72 hr after infection with AdhLAL there was a dose-dependent correction of the severe lipid storage phenotype of Wolman disease fibroblasts. Electron microscopy confirmed significant correction of the lysosomal lipid storage in AdhLAL-infected Wolman disease fibroblasts at the ultrastructural level. Intravenous injection of AdhLAL into wild-type mice resulted in a 13.5-fold increase in hepatic LAL activity, and overexpression of LAL was not associated with toxic side effects. These data demonstrate high-level lysosomal expression of recombinant LAL in vitro and in vivo and show the feasibility of gene therapeutic strategies for the treatment of Wolman disease.
ESTHER : Tietge_2001_Hum.Gene.Ther_12_279
PubMedSearch : Tietge_2001_Hum.Gene.Ther_12_279
PubMedID: 11177564
Gene_locus related to this paper: human-LIPA

Title : Lysosomal acid lipase-deficient mice: depletion of white and brown fat, severe hepatosplenomegaly, and shortened life span - Du_2001_J.Lipid.Res_42_489
Author(s) : Du H , Heur M , Duanmu M , Grabowski GA , Hui DY , Witte DP , Mishra J
Ref : J Lipid Res , 42 :489 , 2001
Abstract : Lysosomal acid lipase (LAL) is essential for the hydrolysis of triglycerides (TG) and cholesteryl esters (CE) in lysosomes. A mouse model created by gene targeting produces no LAL mRNA, protein, or enzyme activity. The lal-/- mice appear normal at birth, survive into adulthood, and are fertile. Massive storage of TG and CE is observed in adult liver, adrenal glands, and small intestine. The age-dependent tissue and gross progression in this mouse model are detailed here. Although lal-/- mice can be bred to give homozygous litters, they die at ages of 7 to 8 months. The lal-/- mice develop enlargement of a single mesenteric lymph node that is full of stored lipids. At 6;-8 months of age, the lal-/- mice have completely absent inguinal, interscapular, and retroperitoneal white adipose tissue. In addition, brown adipose tissue is progressively lost. The plasma free fatty acid levels are significantly higher in lal-/- mice than age-matched lal+/+ mice, and plasma insulin levels were more elevated upon glucose challenge. Energy intake was also higher in lal-/- male mice, although age-matched body weights were not significantly altered from age-matched lal+/+ mice. Early in the disease course, hepatocytes are the main storage cell in the liver; by 3;-8 months, the lipid-stored Kupffer cells progressively fill the liver. The involvement of macrophages throughout the body of lal-/- mice provide evidence for a critical nonappreciated role of LAL in cellular cholesterol and fatty acid metabolism, adipocyte differentiation, and fat mobilization.
ESTHER : Du_2001_J.Lipid.Res_42_489
PubMedSearch : Du_2001_J.Lipid.Res_42_489
PubMedID: 11290820

Title : Enzyme therapy for lysosomal acid lipase deficiency in the mouse - Du_2001_Hum.Mol.Genet_10_1639
Author(s) : Du H , Schiavi S , Levine M , Mishra J , Heur M , Grabowski GA
Ref : Hum Mol Genet , 10 :1639 , 2001
Abstract : Lysosomal acid lipase (LAL) is the critical enzyme for the hydrolysis of the triglycerides (TG) and cholesteryl esters (CE) delivered to lysosomes. Its deficiency produces two human phenotypes, Wolman disease (WD) and cholesteryl ester storage disease (CESD). A targeted disruption of the LAL locus produced a null (lal( -/-)) mouse model that mimics human WD/CESD. The potential for enzyme therapy was tested using mannose terminated human LAL expressed in Pichia pastoris (phLAL), purified, and administered by tail vein injections to lal( -/-) mice. Mannose receptor (MR)-dependent uptake and lysosomal targeting of phLAL were evidenced ex vivo using competitive assays with MR-positive J774E cells, a murine monocyte/macrophage line, immunofluorescence and western blots. Following (bolus) IV injection, phLAL was detected in Kupffer cells, lung macrophages and intestinal macrophages in lal( -/-) mice. Two-month-old lal( -/-) mice received phLAL (1.5 U/dose) or saline injections once every 3 days for 30 days (10 doses). The treated lal( -/-) mice showed nearly complete resolution of hepatic yellow coloration; hepatic weight decreased by approximately 36% compared to PBS-treated lal( -/-) mice. Histologic analyses of numerous tissues from phLAL-treated mice showed reductions in macrophage lipid storage. TG and cholesterol levels decreased by approximately 50% in liver, 69% in spleen and 50% in small intestine. These studies provide feasibility for LAL enzyme therapy in human WD and CESD.
ESTHER : Du_2001_Hum.Mol.Genet_10_1639
PubMedSearch : Du_2001_Hum.Mol.Genet_10_1639
PubMedID: 11487567
Gene_locus related to this paper: mouse-1llip

Title : Sequence and analysis of chromosome 5 of the plant Arabidopsis thaliana - Tabata_2000_Nature_408_823
Author(s) : Tabata S , Kaneko T , Nakamura Y , Kotani H , Kato T , Asamizu E , Miyajima N , Sasamoto S , Kimura T , Hosouchi T , Kawashima K , Kohara M , Matsumoto M , Matsuno A , Muraki A , Nakayama S , Nakazaki N , Naruo K , Okumura S , Shinpo S , Takeuchi C , Wada T , Watanabe A , Yamada M , Yasuda M , Sato S , de la Bastide M , Huang E , Spiegel L , Gnoj L , O'Shaughnessy A , Preston R , Habermann K , Murray J , Johnson D , Rohlfing T , Nelson J , Stoneking T , Pepin K , Spieth J , Sekhon M , Armstrong J , Becker M , Belter E , Cordum H , Cordes M , Courtney L , Courtney W , Dante M , Du H , Edwards J , Fryman J , Haakensen B , Lamar E , Latreille P , Leonard S , Meyer R , Mulvaney E , Ozersky P , Riley A , Strowmatt C , Wagner-McPherson C , Wollam A , Yoakum M , Bell M , Dedhia N , Parnell L , Shah R , Rodriguez M , See LH , Vil D , Baker J , Kirchoff K , Toth K , King L , Bahret A , Miller B , Marra M , Martienssen R , McCombie WR , Wilson RK , Murphy G , Bancroft I , Volckaert G , Wambutt R , Dusterhoft A , Stiekema W , Pohl T , Entian KD , Terryn N , Hartley N , Bent E , Johnson S , Langham SA , McCullagh B , Robben J , Grymonprez B , Zimmermann W , Ramsperger U , Wedler H , Balke K , Wedler E , Peters S , van Staveren M , Dirkse W , Mooijman P , Lankhorst RK , Weitzenegger T , Bothe G , Rose M , Hauf J , Berneiser S , Hempel S , Feldpausch M , Lamberth S , Villarroel R , Gielen J , Ardiles W , Bents O , Lemcke K , Kolesov G , Mayer K , Rudd S , Schoof H , Schueller C , Zaccaria P , Mewes HW , Bevan M , Fransz P
Ref : Nature , 408 :823 , 2000
Abstract : The genome of the model plant Arabidopsis thaliana has been sequenced by an international collaboration, The Arabidopsis Genome Initiative. Here we report the complete sequence of chromosome 5. This chromosome is 26 megabases long; it is the second largest Arabidopsis chromosome and represents 21% of the sequenced regions of the genome. The sequence of chromosomes 2 and 4 have been reported previously and that of chromosomes 1 and 3, together with an analysis of the complete genome sequence, are reported in this issue. Analysis of the sequence of chromosome 5 yields further insights into centromere structure and the sequence determinants of heterochromatin condensation. The 5,874 genes encoded on chromosome 5 reveal several new functions in plants, and the patterns of gene organization provide insights into the mechanisms and extent of genome evolution in plants.
ESTHER : Tabata_2000_Nature_408_823
PubMedSearch : Tabata_2000_Nature_408_823
PubMedID: 11130714
Gene_locus related to this paper: arath-At5g11650 , arath-At5g16120 , arath-at5g18630 , arath-AT5G20520 , arath-At5g21950 , arath-AT5G27320 , arath-CXE15 , arath-F1N13.220 , arath-F14F8.240 , arath-q3e9e4 , arath-q8lae9 , arath-Q8LFB7 , arath-q9ffg7 , arath-q9fij5 , arath-Q9LVU7 , arath-q66gm8 , arath-SCPL34 , arath-B9DFR3 , arath-a0a1p8bcz0

Title : Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana - Mayer_1999_Nature_402_769
Author(s) : Mayer K , Schuller C , Wambutt R , Murphy G , Volckaert G , Pohl T , Dusterhoft A , Stiekema W , Entian KD , Terryn N , Harris B , Ansorge W , Brandt P , Grivell L , Rieger M , Weichselgartner M , de Simone V , Obermaier B , Mache R , Muller M , Kreis M , Delseny M , Puigdomenech P , Watson M , Schmidtheini T , Reichert B , Portatelle D , Perez-Alonso M , Boutry M , Bancroft I , Vos P , Hoheisel J , Zimmermann W , Wedler H , Ridley P , Langham SA , McCullagh B , Bilham L , Robben J , Van der Schueren J , Grymonprez B , Chuang YJ , Vandenbussche F , Braeken M , Weltjens I , Voet M , Bastiaens I , Aert R , Defoor E , Weitzenegger T , Bothe G , Ramsperger U , Hilbert H , Braun M , Holzer E , Brandt A , Peters S , van Staveren M , Dirske W , Mooijman P , Klein Lankhorst R , Rose M , Hauf J , Kotter P , Berneiser S , Hempel S , Feldpausch M , Lamberth S , Van den Daele H , De Keyser A , Buysshaert C , Gielen J , Villarroel R , De Clercq R , van Montagu M , Rogers J , Cronin A , Quail M , Bray-Allen S , Clark L , Doggett J , Hall S , Kay M , Lennard N , McLay K , Mayes R , Pettett A , Rajandream MA , Lyne M , Benes V , Rechmann S , Borkova D , Blocker H , Scharfe M , Grimm M , Lohnert TH , Dose S , de Haan M , Maarse A , Schafer M , Muller-Auer S , Gabel C , Fuchs M , Fartmann B , Granderath K , Dauner D , Herzl A , Neumann S , Argiriou A , Vitale D , Liguori R , Piravandi E , Massenet O , Quigley F , Clabauld G , Mundlein A , Felber R , Schnabl S , Hiller R , Schmidt W , Lecharny A , Aubourg S , Chefdor F , Cooke R , Berger C , Montfort A , Casacuberta E , Gibbons T , Weber N , Vandenbol M , Bargues M , Terol J , Torres A , Perez-Perez A , Purnelle B , Bent E , Johnson S , Tacon D , Jesse T , Heijnen L , Schwarz S , Scholler P , Heber S , Francs P , Bielke C , Frishman D , Haase D , Lemcke K , Mewes HW , Stocker S , Zaccaria P , Bevan M , Wilson RK , de la Bastide M , Habermann K , Parnell L , Dedhia N , Gnoj L , Schutz K , Huang E , Spiegel L , Sehkon M , Murray J , Sheet P , Cordes M , Abu-Threideh J , Stoneking T , Kalicki J , Graves T , Harmon G , Edwards J , Latreille P , Courtney L , Cloud J , Abbott A , Scott K , Johnson D , Minx P , Bentley D , Fulton B , Miller N , Greco T , Kemp K , Kramer J , Fulton L , Mardis E , Dante M , Pepin K , Hillier L , Nelson J , Spieth J , Ryan E , Andrews S , Geisel C , Layman D , Du H , Ali J , Berghoff A , Jones K , Drone K , Cotton M , Joshu C , Antonoiu B , Zidanic M , Strong C , Sun H , Lamar B , Yordan C , Ma P , Zhong J , Preston R , Vil D , Shekher M , Matero A , Shah R , Swaby IK , O'Shaughnessy A , Rodriguez M , Hoffmann J , Till S , Granat S , Shohdy N , Hasegawa A , Hameed A , Lodhi M , Johnson A , Chen E , Marra M , Martienssen R , McCombie WR
Ref : Nature , 402 :769 , 1999
Abstract : The higher plant Arabidopsis thaliana (Arabidopsis) is an important model for identifying plant genes and determining their function. To assist biological investigations and to define chromosome structure, a coordinated effort to sequence the Arabidopsis genome was initiated in late 1996. Here we report one of the first milestones of this project, the sequence of chromosome 4. Analysis of 17.38 megabases of unique sequence, representing about 17% of the genome, reveals 3,744 protein coding genes, 81 transfer RNAs and numerous repeat elements. Heterochromatic regions surrounding the putative centromere, which has not yet been completely sequenced, are characterized by an increased frequency of a variety of repeats, new repeats, reduced recombination, lowered gene density and lowered gene expression. Roughly 60% of the predicted protein-coding genes have been functionally characterized on the basis of their homology to known genes. Many genes encode predicted proteins that are homologous to human and Caenorhabditis elegans proteins.
ESTHER : Mayer_1999_Nature_402_769
PubMedSearch : Mayer_1999_Nature_402_769
PubMedID: 10617198
Gene_locus related to this paper: arath-AT4G00500 , arath-AT4G16690 , arath-AT4G17480 , arath-AT4G24380 , arath-AT4g30610 , arath-o65513 , arath-o65713 , arath-LPAAT , arath-f4jt64

Title : Unique cellular events occurring during the initial interaction of macrophages with matrix-retained or methylated aggregated low density lipoprotein (LDL). Prolonged cell-surface contact during which ldl-cholesteryl ester hydrolysis exceeds ldl protein degradation - Buton_1999_J.Biol.Chem_274_32112
Author(s) : Buton X , Mamdouh Z , Ghosh R , Du H , Kuriakose G , Beatini N , Grabowski GA , Maxfield FR , Tabas I
Ref : Journal of Biological Chemistry , 274 :32112 , 1999
Abstract : A critical event in atherogenesis is the interaction of arterial wall macrophages with subendothelial lipoproteins. Although most studies have investigated this interaction by incubating cultured macrophages with monomeric lipoproteins dissolved in media, arterial wall macrophages encounter lipoproteins that are mostly bound to subendothelial extracellular matrix, and these lipoproteins are often aggregated or fused. Herein, we utilize a specialized cell-culture system to study the initial interaction of macrophages with aggregated low density lipoprotein (LDL) bound to extracellular matrix. The aggregated LDL remains extracellular for a relatively prolonged period of time and becomes lodged in invaginations in the surface of the macrophages. As expected, the degradation of the protein moiety of the LDL was very slow. Remarkably, however, hydrolysis of the cholesteryl ester (CE) moiety of the LDL was 3-7-fold higher than that of the protein moiety, in stark contrast to the situation with receptor-mediated endocytosis of acetyl-LDL. Similar results were obtained using another experimental system in which the degradation of aggregated LDL protein was delayed by LDL methylation rather than by retention on matrix. Additional experiments indicated the following properties of this interaction: (a) LDL-CE hydrolysis is catalyzed by lysosomal acid lipase; (b) neither scavenger receptors nor the LDL receptor appear necessary for the excess LDL-CE hydrolysis; and (c) LDL-CE hydrolysis in this system is resistant to cellular potassium depletion, which further distinguishes this process from receptor-mediated endocytosis. In summary, experimental systems specifically designed to mimic the in vivo interaction of arterial wall macrophages with subendothelial lipoproteins have demonstrated an initial period of prolonged cell-surface contact in which CE hydrolysis exceeds protein degradation.
ESTHER : Buton_1999_J.Biol.Chem_274_32112
PubMedSearch : Buton_1999_J.Biol.Chem_274_32112
PubMedID: 10542246

Title : Mouse lysosomal acid lipase: characterization of the gene and analysis of promoter activity - Du_1998_Gene_208_285
Author(s) : Du H , Duanmu M , Rosa LR
Ref : Gene , 208 :285 , 1998
Abstract : Lysosomal acid lipase (LAL) is required for the hydrolysis of intracellular cholesteryl esters and triglycerides that are delivered to lysosomes by low density lipoprotein (LDL) receptor-mediated endocytosis. To understand that the expression of LAL mRNA and protein is tissue and cell specifically regulated in mice, genomic clones for the mouse lysosomal acid lipase (mLAL) gene were isolated and characterized. The 6.8 kb of the mLAL gene 5'-flanking region was sequenced. Comparisons of mouse and human LAL genes organization revealed identical intron/exon boundaries, except for intron 1 of the mouse gene, and identical exonic length of exons 3-9. The transcription start sites and exon 1 of mLAL were characterized by 5'-RACE-PCR and S1 nuclease mapping. Transfection of 5' flanking deletions of mLAL luciferase reporter gene construct identified positive and negative regulatory elements that varied with cell type. Transfection of three progressively smaller pieces of intron 1 inserted into an SV40 promoter and luciferase reporter gene revealed an enhancer-like activity in intron 1 that is also cell type specific. These studies provide insight into the basis for regulation of this critical enzyme in lipid metabolism.
ESTHER : Du_1998_Gene_208_285
PubMedSearch : Du_1998_Gene_208_285
PubMedID: 98201625

Title : Molecular and enzymatic analyses of lysosomal acid lipase in cholesteryl ester storage disease - Du_1998_Mol.Genet.Metab_64_126
Author(s) : Du H , Sheriff S , Bezerra J , Leonova T , Grabowski GA
Ref : Mol Genet Metab , 64 :126 , 1998
Abstract : Human lysosomal acid lipase (hLAL) is essential for the hydrolysis of cholesteryl esters and triglycerides in the lysosome. Defective hLAL activity leads to two autosomal recessive traits, Wolman disease (WD) or cholesteryl ester storage disease (CESD). Phenotypically, WD has accumulation of both triglycerides and cholesteryl esters, while CESD has mainly elevated cholesteryl esters. We characterized mutations in the hLAL gene from two CESD siblings. By reverse transcriptase-PCR (RT-PCR) and cDNA cloning and sequencing, we identified homozygous deletion mutations of nucleotides 863 to 934, in the hLAL transcript. Normal levels of LAL mRNA were detected. The deletion in mRNA is due to a G to A transition in the last nucleotide of exon 8 of the hLAL gene, a splice junction mutation (E8SJM) that resulted in exon skipping, and a predicted in-frame deletion of the 24 amino acids. [35S]Met metabolic labeling studies in fibroblasts showed a low level of E8SJM LAL ( approximately 38%) that was highly unstable. Heterologous expression of E8SJM LAL in insect cells gave an LAL with low catalytic activity toward cholesteryl oleate and triolein. The effects of this mutation are complex with the production of decreased amounts of an unstable LAL that is catalytically defective. The results suggest that E8SJM leads to essentially a null allele and that the differences in WD and CESD phenotype involve other factors.
ESTHER : Du_1998_Mol.Genet.Metab_64_126
PubMedSearch : Du_1998_Mol.Genet.Metab_64_126
PubMedID: 9705237

Title : Targeted disruption of the mouse lysosomal acid lipase gene: long-term survival with massive cholesteryl ester and triglyceride storage - Du_1998_Hum.Mol.Genet_7_1347
Author(s) : Du H , Duanmu M , Witte D , Grabowski GA
Ref : Hum Mol Genet , 7 :1347 , 1998
Abstract : Lysosomal acid lipase (LAL) is essential for the hydrolysis of the triglycerides and cholesteryl esters in lysosomes. Its deficiency produces two phenotypes, a severe infantile-onset variant, Wolman disease (WD), and a later onset variant, cholesteryl ester storage disease (CESD). A mouse model with a LAL null mutation was produced by targeting disruption of the mouse gene. Homozygote knockout mice (lal -/lal-) produce no LAL mRNA, protein or enzyme activity. The lal-/lal- mice are born in Mendelian ratios, are normal appearing at birth, and follow normal development into adulthood. However, massive accumulation of triglycerides and cholesteryl esters occurs in several organs. By 21 days, the liver develops a yellow-orange color and is approximately 1.5-2.0x larger than normal. The accumulated cholesteryl esters and triglycerides are approximately 30-fold greater than normal. The lal+/lal- mice have approximately 50% of normal LAL activity and do not show lipid accumulation. Male and female lal-/lal- mice are fertile and can be bred to produce progeny. This mouse model is a phenotypic model of human CESD, and a biochemical and histopathologic mimic of human WD. The lal-/lal- mice provide a model to determine the role of LAL in lipid metabolism and the pathogenesis of its deficiency states.
ESTHER : Du_1998_Hum.Mol.Genet_7_1347
PubMedSearch : Du_1998_Hum.Mol.Genet_7_1347
PubMedID: 9700186
Gene_locus related to this paper: mouse-1llip

Title : Tissue and cellular specific expression of murine lysosomal acid lipase mRNA and protein - Du_1996_J.Lipid.Res_37_937
Author(s) : Du H , Witte DP , Grabowski GA
Ref : J Lipid Res , 37 :937 , 1996
Abstract : Lysosomal acid lipase (LAL) is essential to the intracellular control of cholesterol and triglyceride catabolism via the low density lipoprotein (LDL) delivery of these neutral lipids to the lysosome. Deficiency of LAL in humans leads to Wolman disease and cholesteryl ester storage disease that result, respectively, in the intralysosomal storage of both neutral lipids or only cholesteryl esters. The mouse and human LAL cDNAs were cloned. The deduced amino acid sequences from the mouse and human LAL had high similarity (95%) and identity (75%) including conservation of the active center motifs (G-X-S-X-G) and five potential N-glycosylation consensus sequences. Tissue specific expression of LAL mRNA and protein in mouse tissues was evaluated by in situ hybridization and immunofluorescence staining, respectively. The LAL mRNA was expressed at low levels in most tissues. High level expression was found in hepatocytes and splenic and thymic cells. Very high level expression was observed in cells of the small intestinal villi, the zona fasciculata and reticularis of the adrenal cortex, pancreatic acini, and renal tubular epithelium. Significant levels of expression were detected in epithelial cells of choroid plexus in developing mouse embryo by day 12, in liver and lung by day 14, and in small intestine and kidney by day 16. Similar distribution of LAL protein was observed by immunofluorescence stain. Our results show that the expression of LAL is regulated in a tissue- and cell-specific manner that corresponds to the pathologic involvement in Wolman disease.-Du, H., D. P. Witte, and G. A. Grabowski. Tissue and cellular specific expression of murine lysosomal acid lipase mRNA and protein.
ESTHER : Du_1996_J.Lipid.Res_37_937
PubMedSearch : Du_1996_J.Lipid.Res_37_937
PubMedID: 8725147
Gene_locus related to this paper: human-LIPA , mouse-1llip

Title : Characterization of lysosomal acid lipase by site-directed mutagenesis and heterologous expression - Sheriff_1995_J.Biol.Chem_270_27766
Author(s) : Sheriff S , Du H , Grabowski GA
Ref : Journal of Biological Chemistry , 270 :27766 , 1995
Abstract : Lysosomal acid lipase (LAL) is essential for the hydrolysis of cholesterol esters and triglycerides that are delivered to the lysosomes via the low density lipoprotein receptor system. The deficiency of LAL is associated with cholesteryl ester storage disease (CESD) and Wolman's disease (WD). We cloned the human LAL cDNA and expressed the active enzyme in the baculovirus system. Two molecular forms (M(r) approximately 41,000 and approximately 46,000) with different glycosylation were found intracellularly, and approximately 24% of the M(r) approximately 46,000 form was secreted into the medium. Tunicamycin treatment produced only an inactive M(r) approximately 41,000 form. This result implicates glycosylation occupancy in the proper folding for active-site function. Catalytic activity was greater toward cis- than trans-unsaturated fatty acid esters of 4-methylumbelliferone and toward esters with 7-carbon length acyl chains. LAL cleaved cholesterol esters and mono-, tri-, and diglycerides. Heparin had a biphasic effect on enzymatic activity with initial activation followed by inhibition. Inhibition of LAL activity by tetrahydrolipstatin and diethyl p-nitrophenyl phosphate suggested the presence of active serines in binding/catalytic domain(s) of the protein. Site-directed mutagenesis at two putative active centers, GXSXG, showed that Ser153 was important to catalytic activity, whereas Ser99 was not and neither was the catalytic nucleophile. Three reported mutations (L179P, L336P, and delta AG302 deletion) from CESD patients were created and expressed in the Sf9 cell system. None cleaved cholesterol esters, and L179P and L336P cleaved only triolein at approximately 4% of wild-type levels. These results suggest that mechanisms, in addition to LAL defects, may operate in the selective accumulation of cholesterol esters or triglycerides in CESD and WD patients.
ESTHER : Sheriff_1995_J.Biol.Chem_270_27766
PubMedSearch : Sheriff_1995_J.Biol.Chem_270_27766
PubMedID: 7499245
Gene_locus related to this paper: human-LIPA