Kim C

References (13)

Title : Key Metabolic Enzymes Involved in Remdesivir Activation in Human Lung Cells - Li_2021_Antimicrob.Agents.Chemother__AAC0060221
Author(s) : Li R , Liclican A , Xu Y , Pitts J , Niu C , Zhang J , Kim C , Zhao X , Soohoo D , Babusis D , Yue Q , Ma B , Murray BP , Subramanian R , Xie X , Zou J , Bilello JP , Li L , Schultz BE , Sakowicz R , Smith BJ , Shi PY , Murakami E , Feng JY
Ref : Antimicrobial Agents & Chemotherapy , :AAC0060221 , 2021
Abstract : Remdesivir (RDV; GS-5734; Veklury(a)), the first FDA-approved antiviral to treat COVID-19, is a single diastereomer monophosphoramidate prodrug of an adenosine analogue. RDV is taken up in the target cells and metabolized in multiple steps to form the active nucleoside triphosphate (TP) (GS-443902), which in turn acts as a potent and selective inhibitor of multiple viral RNA polymerases. In this report, we profiled the key enzymes involved in the RDV metabolic pathway with multiple parallel approaches: (1) bioinformatic analysis of nucleoside/tide metabolic enzyme mRNA expression using public human tissue and lung single-cell RNAseq datasets; (2) protein and mRNA quantification of enzymes in human lung tissue and primary lung cells; (3) biochemical studies on the catalytic rate of key enzymes; (4) effects of specific enzyme inhibitors on the GS-443902 formation; and (5) the effects of these inhibitors on RDV antiviral activity against SARS-CoV-2 in cell culture. Our data collectively demonstrated that carboxylesterase 1 (CES1) and cathepsin A (CatA) are enzymes involved in hydrolyzing RDV to its alanine intermediate Met X, which is further hydrolyzed to the monophosphate form by histidine triad nucleotide-binding protein 1 (HINT1). The monophosphate is then consecutively phosphorylated to diphosphate and triphosphate by cellular phosphotransferases. Our data support the hypothesis that the unique properties of RDV prodrug not only allow lung-specific accumulation critical for the treatment of respiratory viral infection such as COVID-19, they also enable efficient intracellular metabolism of RDV and its Met X to monophosphate and successive phosphorylation to form the active TP in disease-relevant cells.
ESTHER : Li_2021_Antimicrob.Agents.Chemother__AAC0060221
PubMedSearch : Li_2021_Antimicrob.Agents.Chemother__AAC0060221
PubMedID: 34125594

Title : Simultaneous Time-concentration Analysis of Soman and VX Adducts to Butyrylcholinesterase and Albumin by LC-MS-MS - Lee_2018_J.Anal.Toxicol_42_293
Author(s) : Lee JY , Kim C , Lee YH
Ref : J Anal Toxicol , 42 :293 , 2018
Abstract : A sensitive method for the purification and determination of two protein adducts, organophosphorus (OP)-BChE and OP-albumin adducts, in a single sample using a simultaneous sample preparation method was developed and validated using liquid chromatography-tandem mass spectrometry. First, we isolated O-ethyl S-2-diisopropylaminoethyl methyl phosphonothiolate (VX) and O-pinacolyl methylphosphonofluoridate (soman, GD)-BChE adducts using an immunomagnetic separation (IMS) method and the HiTrap Blue affinity column was subsequently used to isolate and purify VX and GD-albumin adducts from the plasma of rhesus monkeys exposed to nerve agents. Additionally, we examined the time-concentration profiles of two biomarkers, VX and GD-nonapeptides and VX and GD-tyrosines, derived from OP-BChE and OP-albumin adducts up to 8 weeks after exposure. Based on the results, we determined that VX and GD-tyrosine is more suitable than VX and GD-nonapeptide as a biomarker owing to its longevity. This integrated approach is expected to be applicable for the quantification of other OP-BChE and OP-albumin adducts in human plasma, thus serving as a potential generic assay for exposure to nerve agents.
ESTHER : Lee_2018_J.Anal.Toxicol_42_293
PubMedSearch : Lee_2018_J.Anal.Toxicol_42_293
PubMedID: 29618078

Title : Fluorogenic structure activity library pinpoints molecular variations in substrate specificity of structurally homologous esterases - White_2018_J.Biol.Chem_293_13851
Author(s) : White A , Koelper A , Russell A , Larsen EM , Kim C , Lavis LD , Hoops GC , Johnson RJ
Ref : Journal of Biological Chemistry , 293 :13851 , 2018
Abstract : Cellular esterases catalyze many essential biological functions by performing hydrolysis reactions on diverse substrates. The promiscuity of esterases complicates assignment of their substrate preferences and biological functions. To identify universal factors controlling esterase substrate recognition, we designed a 32-member structure-activity relationship (SAR) library of fluorogenic ester substrates and used this library to systematically interrogate esterase preference for chain length, branching patterns, and polarity to differentiate common classes of esterase substrates. Two structurally homologous bacterial esterases were screened against this library, refining their previously broad overlapping substrate specificity. Vibrio cholerae esterase ybfF displayed a preference for gamma-position thioethers and ethers, whereas Rv0045c from Mycobacterium tuberculosis displayed a preference for branched substrates with and without thioethers. We determined that this substrate differentiation was partially controlled by individual substrate selectivity residues Tyr-119 in ybfF and His-187 in Rv0045c; reciprocal substitution of these residues shifted each esterase's substrate preference. This work demonstrates that the selectivity of esterases is tuned based on transition state stabilization, identifies thioethers as an underutilized functional group for esterase substrates, and provides a rapid method for differentiating structural isozymes. This SAR library could have multifaceted future applications, including in vivo imaging, biocatalyst screening, molecular fingerprinting, and inhibitor design.
ESTHER : White_2018_J.Biol.Chem_293_13851
PubMedSearch : White_2018_J.Biol.Chem_293_13851
PubMedID: 30006352
Gene_locus related to this paper: myctu-RV0045C , vibch-VC2097

Title : Measuring the Global Substrate Specificity of Mycobacterial Serine Hydrolases Using a Library of Fluorogenic Ester Substrates - Bassett_2018_ACS.Infect.Dis_4_904
Author(s) : Bassett B , Waibel B , White A , Hansen H , Stephens D , Koelper A , Larsen EM , Kim C , Glanzer A , Lavis LD , Hoops GC , Johnson RJ
Ref : ACS Infect Dis , 4 :904 , 2018
Abstract : Among the proteins required for lipid metabolism in Mycobacterium tuberculosis are a significant number of uncharacterized serine hydrolases, especially lipases and esterases. Using a streamlined synthetic method, a library of immolative fluorogenic ester substrates was expanded to better represent the natural lipidomic diversity of Mycobacterium. This expanded fluorogenic library was then used to rapidly characterize the global structure activity relationship (SAR) of mycobacterial serine hydrolases in M. smegmatis under different growth conditions. Confirmation of fluorogenic substrate activation by mycobacterial serine hydrolases was performed using nonspecific serine hydrolase inhibitors and reinforced the biological significance of the SAR. The hydrolases responsible for the global SAR were then assigned using gel-resolved activity measurements, and these assignments were used to rapidly identify the relative substrate specificity of previously uncharacterized mycobacterial hydrolases. These measurements provide a global SAR of mycobacterial hydrolase activity, a picture of cycling hydrolase activity, and a detailed substrate specificity profile for previously uncharacterized hydrolases.
ESTHER : Bassett_2018_ACS.Infect.Dis_4_904
PubMedSearch : Bassett_2018_ACS.Infect.Dis_4_904
PubMedID: 29648787

Title : CHRNA5 risk variant predicts delayed smoking cessation and earlier lung cancer diagnosis--a meta-analysis - Chen_2015_J.Natl.Cancer.Inst_107_
Author(s) : Chen LS , Hung RJ , Baker T , Horton A , Culverhouse R , Saccone N , Cheng I , Deng B , Han Y , Hansen HM , Horsman J , Kim C , Lutz S , Rosenberger A , Aben KK , Andrew AS , Breslau N , Chang SC , Dieffenbach AK , Dienemann H , Frederiksen B , Han J , Hatsukami DK , Johnson EO , Pande M , Wrensch MR , McLaughlin J , Skaug V , van der Heijden HF , Wampfler J , Wenzlaff A , Woll P , Zienolddiny S , Bickeboller H , Brenner H , Duell EJ , Haugen A , Heinrich J , Hokanson JE , Hunter DJ , Kiemeney LA , Lazarus P , Le Marchand L , Liu G , Mayordomo J , Risch A , Schwartz AG , Teare D , Wu X , Wiencke JK , Yang P , Zhang ZF , Spitz MR , Kraft P , Amos CI , Bierut LJ
Ref : J Natl Cancer Inst , 107 : , 2015
Abstract : BACKGROUND: Recent meta-analyses show strong evidence of associations among genetic variants in CHRNA5 on chromosome 15q25, smoking quantity, and lung cancer. This meta-analysis tests whether the CHRNA5 variant rs16969968 predicts age of smoking cessation and age of lung cancer diagnosis.
METHODS: Meta-analyses examined associations between rs16969968, age of quitting smoking, and age of lung cancer diagnosis in 24 studies of European ancestry (n = 29 072). In each dataset, we used Cox regression models to evaluate the association between rs16969968 and the two primary phenotypes (age of smoking cessation among ever smokers and age of lung cancer diagnosis among lung cancer case patients) and the secondary phenotype of smoking duration. Heterogeneity across studies was assessed with the Cochran Q test. All statistical tests were two-sided.
RESULTS: The rs16969968 allele (A) was associated with a lower likelihood of smoking cessation (hazard ratio [HR] = 0.95, 95% confidence interval [CI] = 0.91 to 0.98, P = .0042), and the AA genotype was associated with a four-year delay in median age of quitting compared with the GG genotype. Among smokers with lung cancer diagnoses, the rs16969968 genotype (AA) was associated with a four-year earlier median age of diagnosis compared with the low-risk genotype (GG) (HR = 1.08, 95% CI = 1.04 to 1.12, P = 1.1*10(-5)). CONCLUSION: These data support the clinical significance of the CHRNA5 variant rs16969968. It predicts delayed smoking cessation and an earlier age of lung cancer diagnosis in this meta-analysis. Given the existing evidence that this CHRNA5 variant predicts favorable response to cessation pharmacotherapy, these findings underscore the potential clinical and public health importance of rs16969968 in CHRNA5 in relation to smoking cessation success and lung cancer risk.
ESTHER : Chen_2015_J.Natl.Cancer.Inst_107_
PubMedSearch : Chen_2015_J.Natl.Cancer.Inst_107_
PubMedID: 25873736

Title : Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template - Grimster_2012_J.Am.Chem.Soc_134_6732
Author(s) : Grimster NP , Stump B , Fotsing JR , Weide T , Talley TT , Yamauchi JG , Nemecz A , Kim C , Ho KY , Sharpless KB , Taylor P , Fokin VV
Ref : Journal of the American Chemical Society , 134 :6732 , 2012
Abstract : Nicotinic acetylcholine receptors (nAChRs), which are responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nAChRs are pentameric, composed of various permutations of alpha (alpha2 to alpha10) and beta (beta2 to beta4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates the development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building-block azides and alkynes to form stable 1,2,3-triazoles was used to generate the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product for the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at the subunit interfaces of an oligomeric protein and can thus be used as a tool for identifying novel candidate nAChR ligands. The crystal structure of one of the in situ-formed triazole-AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.
ESTHER : Grimster_2012_J.Am.Chem.Soc_134_6732
PubMedSearch : Grimster_2012_J.Am.Chem.Soc_134_6732
PubMedID: 22394239

Title : Pharmacokinetic comparison of orally disintegrating and conventional donepezil formulations in healthy Korean male subjects: a single-dose, randomized, open-label, 2-sequence, 2-period crossover study - Kim_2011_Clin.Ther_33_965
Author(s) : Kim KA , Lim JL , Kim C , Park JY
Ref : Clin Ther , 33 :965 , 2011
Abstract : BACKGROUND: Donepezil is a potent inhibitor of acetylcholinesterase, an enzyme that is targeted in the treatment of Alzheimer's disease. OBJECTIVE: The purpose of this study was to compare the pharmacokinetic characteristics of orally disintegrating (test) and conventional (reference) donepezil formulations to satisfy the regulatory requirement for marketing. METHODS: A single-center randomized, single-dose, open-label, 2-way crossover study with a 21-day washout period was conducted in 22 healthy volunteers. Plasma samples for the analysis of donepezil were collected up to 240 hours after drug administration. Participants received either reference or test drug formulation of 10 mg donepezil in the first period and the alternative formulation in the second period. Plasma concentrations of donepezil were determined by validated high-performance liquid chromatography coupled to tandem mass spectrometry detection. Pharmacokinetic parameters, including C(max) and AUC, were determined by noncompartmental analysis. ANOVA was carried out using log-transformed C(max) and AUC, and the mean ratios and their 90% CIs were calculated. The safety profiles and tolerabilities of the 2 formulations were also assessed based on laboratory tests, 12-lead ECGs, vital signs, and physical examinations. RESULTS: Of the 22 participants initially enrolled, 18 healthy Korean participants completed both treatment periods. Four subjects did not complete both treatments: 3 subjects withdrew consent for personal reasons, and 1 subject experienced adverse events. No significant differences in pharmacokinetic parameters between the 2 formulations were observed. The mean (SD) age, height, and weight of the participants were 25.8 (4.1) years, 173.6 (5.7) cm, and 68.9 (7.8) kg, respectively. The mean (SD) C(max), AUC(last), and AUC(inf) for the reference formulation were 33.26 (6.58) ng/mL, 1521.69 (344.04) ng x h/mL, and 1691.46 (443.05) ng x h/mL, respectively. Corresponding values for the test formulation were 34.23 (6.79) ng/mL, 1554.33 (390.23) ng x h/mL, and 1718.27 (447.03) ng x h/mL, respectively. The median T(max) was 2 hours (range, 1-3 hours) for the reference and test formulations. The geometric mean ratios (90% CI) between the 2 formulations of donepezil were 102.9 (96.8-109.5) for C(max), 102.3 (96.1-108.9) for AUC(last), and 101.6 (95.4-108.2) for AUC(0-infinity), respectively. During the study, 15 and 14 adverse events were reported for the reference and test formulations, respectively, and all were transient, mild, and resolved during the treatment period. These adverse events included 7 cases of nausea, 3 cases of headache, and 1 case each of dizziness, vomiting, chills, and sweating. All adverse events were considered related to the study drugs. CONCLUSION: This study found that the test and reference formulations met the regulatory criteria for pharmacokinetic equivalence in these fasting healthy Korean male subjects. Both donepezil formulations appeared to be generally well tolerated.
ESTHER : Kim_2011_Clin.Ther_33_965
PubMedSearch : Kim_2011_Clin.Ther_33_965
PubMedID: 21723605

Title : Chemoenzymatic synthesis of rivastigmine via dynamic kinetic resolution as a key step - Han_2010_J.Org.Chem_75_3105
Author(s) : Han K , Kim C , Park J , Kim MJ
Ref : J Org Chem , 75 :3105 , 2010
Abstract : A practical and efficient procedure for the synthesis of rivastigmine was developed. This procedure includes dynamic kinetic resolution using a polymer-bound ruthenium complex and a lipase in combination as a key step. Enantiopure (-)-rivastigmine was obtained from commercially available 3'-hydroxyacetophenone via five steps in overall 57% yield.
ESTHER : Han_2010_J.Org.Chem_75_3105
PubMedSearch : Han_2010_J.Org.Chem_75_3105
PubMedID: 20345141

Title : Empirical analysis of transcriptional activity in the Arabidopsis genome - Yamada_2003_Science_302_842
Author(s) : Yamada K , Lim J , Dale JM , Chen H , Shinn P , Palm CJ , Southwick AM , Wu HC , Kim C , Nguyen M , Pham P , Cheuk R , Karlin-Newmann G , Liu SX , Lam B , Sakano H , Wu T , Yu G , Miranda M , Quach HL , Tripp M , Chang CH , Lee JM , Toriumi M , Chan MM , Tang CC , Onodera CS , Deng JM , Akiyama K , Ansari Y , Arakawa T , Banh J , Banno F , Bowser L , Brooks S , Carninci P , Chao Q , Choy N , Enju A , Goldsmith AD , Gurjal M , Hansen NF , Hayashizaki Y , Johnson-Hopson C , Hsuan VW , Iida K , Karnes M , Khan S , Koesema E , Ishida J , Jiang PX , Jones T , Kawai J , Kamiya A , Meyers C , Nakajima M , Narusaka M , Seki M , Sakurai T , Satou M , Tamse R , Vaysberg M , Wallender EK , Wong C , Yamamura Y , Yuan S , Shinozaki K , Davis RW , Theologis A , Ecker JR
Ref : Science , 302 :842 , 2003
Abstract : Functional analysis of a genome requires accurate gene structure information and a complete gene inventory. A dual experimental strategy was used to verify and correct the initial genome sequence annotation of the reference plant Arabidopsis. Sequencing full-length cDNAs and hybridizations using RNA populations from various tissues to a set of high-density oligonucleotide arrays spanning the entire genome allowed the accurate annotation of thousands of gene structures. We identified 5817 novel transcription units, including a substantial amount of antisense gene transcription, and 40 genes within the genetically defined centromeres. This approach resulted in completion of approximately 30% of the Arabidopsis ORFeome as a resource for global functional experimentation of the plant proteome.
ESTHER : Yamada_2003_Science_302_842
PubMedSearch : Yamada_2003_Science_302_842
PubMedID: 14593172
Gene_locus related to this paper: arath-AT2G42690 , arath-AT4g30610 , arath-At5g13640 , arath-AT5G20520 , arath-AT5G27320 , arath-CGEP , arath-clh1 , arath-clh2 , arath-CXE12 , arath-CXE15 , arath-SCP25 , arath-F14F8.240 , arath-MES6 , arath-LCAT1 , arath-PLA11 , arath-PLA15 , arath-PLA16 , arath-PLA17 , arath-SCP8 , arath-SCP11 , arath-SCP40 , arath-MES14 , arath-AXR4 , arath-SFGH , arath-B9DFR3 , arath-pae2

Title : Cloning and sequence analysis of the estA gene encoding enzyme for producing (R)-beta-acetylmercaptoisobutyric acid from Pseudomonas aeruginosa 1001 - Lee_2000_J.Biosci.Bioeng_90_684
Author(s) : Lee J , Boyapati G , Song K , Rhee S , Kim C
Ref : J Biosci Bioeng , 90 :684 , 2000
Abstract : The estA gene encoding the enzyme that catalyzes the production of (R)-beta-acetylmercaptoisobutyric acid from (R,S)-ester from Pseudomonas aeruginosa 1001, was cloned in Escherichia coli and its nucleotide sequence was determined, revealing the presumed open reading frame encoding a polypeptide of 316 amino acid residues (948 nucleotides). The overall A + T and C + G compositions were 32.59% and 67.41%, respectively. The amino acid sequence of the estA gene product showed a significant similarity with that of the triacylglycerol lipase from Psychrobacter immobilis (38% identity), triacylglycerol lipase from Moraxella sp. (36% identity), and two forms of carboxyl esterases from Acinetobacter calcoaceticus (17% and 17% identities). The deduced amino acid sequences have a pentapeptide consensus sequence, G-X-S-X-G, having an active serine residue, and another active site, dipeptides H-G, located at 70-100 amino acids upstream of the G-X-S-X-G consensus sequence.
ESTHER : Lee_2000_J.Biosci.Bioeng_90_684
PubMedSearch : Lee_2000_J.Biosci.Bioeng_90_684
PubMedID: 16232934
Gene_locus related to this paper: pseae-PA2949

Title : Regulation of proteolytic processing of the amyloid beta-protein precursor of Alzheimer's disease in transfected cell lines and in brain slices - Nitsch_1994_J.Neural.Transm.Suppl_44_21
Author(s) : Nitsch RM , Slack BE , Farber SA , Schulz JG , Deng M , Kim C , Borghesani PR , Korver W , Wurtman RJ , Growdon JH
Ref : J Neural Transm Suppl , 44 :21 , 1994
Abstract : beta A4 is the principal component of Alzheimer's disease brain amyloid. It is derived from proteolytic processing of amyloid beta-protein precursors (APP), a family of transmembrane glycoproteins. Secretion of APPs, a secreted proteolytic derivative that is cleaved within the beta A4 domain of APP, is increased many-fold by the activation of cell-surface receptors, like the muscarinic m1 and m3 receptor subtypes, which are coupled to protein kinase C. Concomitantly, their activation decreases the formation of both secreted soluble beta A4 and of endosomal-lysosomal C-terminal APP derivatives. These data suggest that muscarinic m1 and m3 receptors accelerate non-amyloidogenic APP processing and depress the formation of potentially amyloidogenic derivatives. Other receptors that stimulate APPs secretion include those for bradykinin, vasopressin, and interleukin-1 receptors. A similar control mechanism is present in rat brain tissue slices, in which the release of both APPs and endogenous neurotransmitters is increased by electrical depolarization. This increase is tetrodotoxin-sensitive and frequency-dependent, suggesting that APPs release may normally depend on neuronal activity. Taken together, our findings suggest that specific receptor agonists might be effective in reducing the formation of potentially amyloidogenic APP derivatives in vivo.
ESTHER : Nitsch_1994_J.Neural.Transm.Suppl_44_21
PubMedSearch : Nitsch_1994_J.Neural.Transm.Suppl_44_21
PubMedID: 7897393

Title : Phosphorylation and desensitization of human m2 muscarinic cholinergic receptors by two isoforms of the beta-adrenergic receptor kinase - Richardson_1993_J.Biol.Chem_268_13650
Author(s) : Richardson RM , Kim C , Benovic JL , Hosey MM
Ref : Journal of Biological Chemistry , 268 :13650 , 1993
Abstract : Studies of the human m2 (hm2) muscarinic cholinergic receptors (mAChR) have been performed to provide further insights into the potential regulation of these receptors by isoforms of the beta-adrenergic receptor kinase (beta ARK). The hm2 mAChR and the isoforms beta ARK1 and beta ARK2 were individually expressed in, and purified from, insect Sf9 cells infected with recombinant baculoviruses. The expressed hm2 receptors were tested as substrates for beta ARK1 and beta ARK2 in vitro using concentrations of receptors and kinases similar to those found in intact cells. The hm2 mAChR were phosphorylated in an agonist-dependent manner to 4-5 mol of phosphate/mol of receptor by beta ARK1 or beta ARK2. The reactions were highly dependent on agonist; the antagonist atropine, and heparin, a beta ARK inhibitor, both prevented the beta ARK-mediated phosphorylation. The rates of phosphorylation catalyzed by both isoforms were similar, with half-maximal phosphorylation occurring in less than 5 min. Under the conditions employed the stoichiometries, but not the rates, of phosphorylation catalyzed by both kinases were increased 2-3-fold by either the heterotrimeric G-protein G(o) or the beta gamma subunits of transducin. Phosphopeptide mapping experiments indicated that similar sites were phosphorylated by the two beta ARK isoforms. In order to test for functional effects of the phosphorylation mediated by the beta ARK isoforms, the receptors were reconstituted with purified G(o) and were tested for their ability to stimulate guanosine 5'-3-O-(thio)triphosphate (GTP gamma S) binding. The conditions leading to maximal receptor phosphorylation resulted in a 30-50% reduction in the ability of the receptors to stimulate GTP gamma S binding to G(o). The results demonstrate that the hm2 mAChR are excellent substrates in vitro for both beta ARK1 and beta ARK2 and that extensive phosphorylation by these enzymes occurs in the presence of the beta gamma subunits of G proteins. The beta ARK-mediated phosphorylation of the m2 mAChR causes a perturbation of receptor/G-protein coupling.
ESTHER : Richardson_1993_J.Biol.Chem_268_13650
PubMedSearch : Richardson_1993_J.Biol.Chem_268_13650
PubMedID: 8514797

Title : Receptor-coupled amyloid precursor protein processing - Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
Author(s) : Nitsch RM , Slack BE , Farber SA , Borghesani PR , Schulz JG , Kim C , Felder CC , Growdon JH , Wurtman RJ
Ref : Annals of the New York Academy of Sciences , 695 :122 , 1993
Abstract : The family of beta-amyloid protein precursors (APP) can be processed via several alternative proteolytic pathways. Some generate potentially amyloidogenic APP derivatives, whereas others preclude the formation of such fragments. The cellular mechanisms regulating the relative activities of these pathways are thus important in determining the factors contributing to the formation of amyloidogenic APP derivatives. In order to investigate whether cell-surface receptor activity can regulate APP processing, HEK 293 cell lines stably expressing human muscarinic acetylcholine receptors (mAChR; subtypes m1, m2, m3, m4) were stimulated with the muscarinic agonist carbachol, and the release of APP derivatives was measured. Carbachol increased the release of large amino-terminal APP-fragments 4- to 6-fold in cell lines expressing the m1 or m3 receptors but not in those expressing m2 or m4 subtypes. This increase was blocked by various protein kinase inhibitors and mimicked by phorbol esters, indicating that it is mediated by protein kinase activation, presumably by protein kinase C (PKC). To determine whether additional cell-surface receptor types linked to this signal transduction pathway could also regulate APP processing, we stimulated differentiated PC-12 cells with bradykinin and found that this neuropeptide also increased the secretion of amino-terminal APP derivatives. We next investigated the possibility that neuronal depolarization might affect APP processing in mammalian brain. Electrically stimulated rat hippocampal slices released two times more amino-terminal APP derivatives than unstimulated control slices. This release increased with increasing stimulation frequencies in the physiological firing range of hippocampal pyramidal cells, and was blocked by tetrodotoxin. These results suggest that, in brain, APP processing is regulated by neuronal activity.
ESTHER : Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
PubMedSearch : Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
PubMedID: 8239269