Zevin-Sonkin D

References (10)

Title : Expression and tissue-specific assembly of human butyrylcholine esterase in microinjected Xenopus laevis oocytes - Soreq_1989_J.Biol.Chem_264_10608
Author(s) : Soreq H , Seidman S , Dreyfus PA , Zevin-Sonkin D , Zakut H
Ref : Journal of Biological Chemistry , 264 :10608 , 1989
Abstract : Cholinesterases represent a ubiquitous, polymorphic family of acetylcholine hydrolyzing enzymes. The multileveled tissue-specific heterogeneity which characterizes these enzymes makes the cholinesterases an appropriate model for studying the mechanisms involved in regulating divergent pathways in protein biogenesis. For this purpose, a cDNA coding for human butyrylcholine esterase (BCHE) was subcloned into the SP 6 transcription vector. Synthetic mRNA transcribed from this construct was microninjected into Xenopus laevis oocytes alone, and in conjunction with poly(A)+ RNAs extracted from human brain or muscle. Injected BCHE-mRNA induced the biosynthesis of a protein exhibiting the catalytic activity, substrate specificity, and sensitivity to selective inhibitors characteristic of native human serum BCHE, and clearly distinct from the related enzyme acetylcholinesterase (AChE). The nascent BCHE was reproducibly distributed into low salt-soluble and detergent-extractable pools. Sucrose gradient analysis demonstrated that the nascent human enzyme was capable of limited subunit assembly, appearing as functional dimeric molecules in both of these fractions. Co-injection with brain or muscle-derived mRNAs facilitated higher order oligomeric assembly. Co-injected brain mRNA induced the appearance of tetramers while co-injected muscle mRNA induced the appearance of an array of heavy molecular forms, including a heavy 16 S form. These results indicate that the molecular determinants which distinguish BCHE from AChE are inherent to its primary amino acid sequence and that additional, tissue-specific protein(s) are involved in the modulation of subunit assembly within particular biological milieues.
ESTHER : Soreq_1989_J.Biol.Chem_264_10608
PubMedSearch : Soreq_1989_J.Biol.Chem_264_10608
PubMedID: 2732242

Title : Cross-homologies and structural differences between human cholinesterases revealed by antibodies against cDNA-produced human butyrylcholinesterase peptides - Dreyfus_1988_J.Neurochem_51_1858
Author(s) : Dreyfus PA , Zevin-Sonkin D , Seidman S , Prody CA , Zisling R , Zakut H , Soreq H
Ref : Journal of Neurochemistry , 51 :1858 , 1988
Abstract : To study the polymorphism of human cholinesterases (ChEs) at the levels of primary sequence and three-dimensional structure, a fragment of human butyrylcholinesterase (BCHE) cDNA was subcloned into the pEX bacterial expression vector and its polypeptide product analyzed. Immunoblot analysis revealed that the clone-produced BCHE peptides interact specifically with antibodies against human and Torpedo acetylcholinesterase (AChE). Rabbit polyclonal antibodies prepared against the purified clone-produced BCHE polypeptides interacted in immunoblots with denatured serum BCHE as well as with purified and denatured erythrocyte AChE. In contrast, native BCHE tetramers from human serum, but not AChE dimers from erythrocytes, interacted with these antibodies in solution to produce antibody-enzyme complexes that could be precipitated by second antibodies and that sedimented faster than the native enzyme in sucrose gradient centrifugation. Furthermore, both AChE and BCHE dimers from muscle extracts, but not BCHE tetramers from muscle, interacted with these antibodies. To reveal further whether the anti-cloned BCHE antibodies would interact in situ with ChEs in the neuromuscular junction, bundles of muscle fibers were microscopically dissected from the region in fetal human diaphragm that is innervated by the phrenic nerve. Muscle fibers incubated with the antibodies and with 125I-Protein A were subjected to emulsion autoradiography, followed by cytochemical ChE staining. The anti-cloned BCHE antibodies, as well as anti-Torpedo AChE antibodies, created patches of silver grains in the muscle endplate region stained for ChE, under conditions where control sera did not. These findings demonstrate that the various forms of human AChE and BCHE in blood and in neuromuscular junctions share sequence homologies, but also display structural differences between distinct molecular forms within particular tissues, as well as between similarly sedimenting molecular forms from different tissues.
ESTHER : Dreyfus_1988_J.Neurochem_51_1858
PubMedSearch : Dreyfus_1988_J.Neurochem_51_1858
PubMedID: 2460589

Title : Human cholinesterase genes localized by hybridization to chromosomes 3 and 16 - Soreq_1987_Hum.Genet_77_325
Author(s) : Soreq H , Zamir R , Zevin-Sonkin D , Zakut H
Ref : Hum Genet , 77 :325 , 1987
Abstract : A cloned human cDNA for cholinesterase (ChE) was used as a probe for in situ hybridization to spread lymphocyte chromosomes to map the structural human CHE genes to distinct chromosomal regions. The recent genetic linkage assignment of the CHE1 locus of the CHE gene to chromosome 3q was confirmed and further refined to 3q21-q26, close to the genes coding for transferrin (TF) and transferrin receptor (TFRC). The CHE1 allele localizes to a 3q region that is commonly mutated and then associated with abnormal megakaryocyte proliferation in acute myelodysplastic anomalies. In view of earlier findings that ChE inhibitors induce megakaryocytopoiesis in culture, this localization may indicate that ChEs are involved in regulating the differentiation of megakaryocytes. A second site for ChEcDNA hybridization was found on chromosome 16p11-q23, demonstrating that the CHE2 locus of the cholinesterase gene, which directs the production of the common C5 variant of serum ChE, also codes for a structural subunit of the enzyme and is localized on the same chromosome with the haptoglobin (HP) gene, both genes being found on the long arm of chromosome 16. The finding of two sites for ChEcDNA hybridization suggests that the two loci coding for human ChEs may include nonidentical sequences responsible for the biochemical differences between ChE variants.
ESTHER : Soreq_1987_Hum.Genet_77_325
PubMedSearch : Soreq_1987_Hum.Genet_77_325
PubMedID: 3692476

Title : Isolation and characterization of full-length cDNA clones coding for cholinesterase from fetal human tissues - Prody_1987_Proc.Natl.Acad.Sci.U.S.A_84_3555
Author(s) : Prody CA , Zevin-Sonkin D , Gnatt A , Goldberg O , Soreq H
Ref : Proceedings of the National Academy of Sciences of the United States of America , 84 :3555 , 1987
Abstract : To study the primary structure and regulation of human cholinesterases, oligodeoxynucleotide probes were prepared according to a consensus peptide sequence present in the active site of both human serum pseudocholinesterase (BtChoEase; EC and Torpedo electric organ "true" acetylcholinesterase (AcChoEase; EC Using these probes, we isolated several cDNA clones from lambda gt10 libraries of fetal brain and liver origins. These include 2.4-kilobase cDNA clones that code for a polypeptide containing a putative signal peptide and the N-terminal, active site, and C-terminal peptides of human BtChoEase, suggesting that they code either for BtChoEase itself or for a very similar but distinct fetal form of cholinesterase. In RNA blots of poly(A)+ RNA from the cholinesterase-producing fetal brain and liver, these cDNAs hybridized with a single 2.5-kilobase band. Blot hybridization to human genomic DNA revealed that these fetal BtChoEase cDNA clones hybridize with DNA fragments of the total length of 17.5 kilobases, and signal intensities indicated that these sequences are not present in many copies. Both the cDNA-encoded protein and its nucleotide sequence display striking homology to parallel sequences published for Torpedo AcChoEase. These findings demonstrate extensive homologies between the fetal BtChoEase encoded by these clones and other cholinesterases of various forms and species.
ESTHER : Prody_1987_Proc.Natl.Acad.Sci.U.S.A_84_3555
PubMedSearch : Prody_1987_Proc.Natl.Acad.Sci.U.S.A_84_3555
PubMedID: 3035536
Gene_locus related to this paper: human-BCHE

Title : Use of synthetic oligodeoxynucleotide probes for the isolation of a human cholinesterase cDNA clone - Prody_1986_J.Neurosci.Res_16_25
Author(s) : Prody CA , Zevin-Sonkin D , Gnatt A , Koch R , Zisling R , Goldberg O , Soreq H
Ref : Journal of Neuroscience Research , 16 :25 , 1986
Abstract : Cholinesterases are serine esterases that rapidly hydrolyze the neurotransmitter acetylcholine. In humans, cholinesterases exhibit extensive polymorphism in terms of their substrate specificity, sensitivity to selective inhibitors, hydrophobicity, and cellular as well as subcellular localization. It is not yet known whether the various cholinesterase forms originate from different genes or are products of posttranscriptional and posttranslational processing. The extent to which these enzyme forms are homologous in their amino acid sequence is also not known. However, a consensus organophosphate-binding hexapeptide sequence Phe-Gly-Glu-Ser-Ala-Gly was found both in "true" acetylcholinesterase from the electric organ of Torpedo [McPhee-Quigley et al: J Biol Chem 260:12185-12189, 1985] and in "pseudocholinesterase" (butyrylcholinesterase) from human serum [Lockridge: "Cholinesterases--Fundamental and Applied Aspects." New York: de Gruyter pp 5-12, 1984], suggesting that this region in the protein is conserved in all cholinesterases. Based on this common sequence, we prepared synthetic oligodeoxynucleotides and used them as labeled probes to screen a cDNA library from fetal human brain mRNA, cloned in lambda gt10 phages. A cDNA clone of 770 nucleotides in length was isolated. It contains an open reading frame terminating with the sequence Ser-Val-Thr-Leu-Phe-Gly-Glu-Ser-Ala-Gly-Ala-Ala, which includes the consensus hexapeptide used for designing the DNA probe. Furthermore, the sequence of this 12-amino acid peptide is identical to the sequence reported for the organophosphate binding site of human serum pseudocholinesterase [Lockridge: "Cholinesterases--Fundamental and Applied Aspects." New York: de Gruyter, pp 5-12, 1984]. These findings confirm that the isolated clone is indeed part of a human cholinesterase cDNA.
ESTHER : Prody_1986_J.Neurosci.Res_16_25
PubMedSearch : Prody_1986_J.Neurosci.Res_16_25
PubMedID: 3755763

Title : The use of mRNA translation in vitro and in ovo followed by crossed immunoelectrophoretic autoradiography to study the biosynthesis of human cholinesterases - Soreq_1986_Cell.Mol.Neurobiol_6_227
Author(s) : Soreq H , Dziegielewska KM , Zevin-Sonkin D , Zakut H
Ref : Cellular Molecular Neurobiology , 6 :227 , 1986
Abstract : The synthesis of various cholinesterases in different fetal human tissues was studied using in vitro and in ovo translation of poly(A)+ RNA, followed by crossed immunoelectrophoretic autoradiography. When unfractionated poly(A)+ mRNA from fetal brain, muscle, or liver was translated in vitro, in the reticulocyte lysate cell-free system, polypeptides were synthesized which reacted with antibodies against either "true" acetylcholinesterase (acetylcholine hydrolase; EC or "pseudo", butyrylcholinesterase (acylcholine acylhydrolase; EC The two nascent cholinesterases could be separated by crossed immunoelectrophoresis followed by autoradiography, suggesting that acetylcholinesterase and butyrylcholinesterase are produced in all three tissues from nascent polypeptides containing different immunological domains. To examine whether the biosynthesis of cholinesterases includes posttranslational processing events, Xenopus oocytes were microinjected with mRNA from these tissues. Immunoelectrophoretic analysis of oocyte intracellular homogenates and incubation medium revealed various precipitation arcs, reflecting the synthesis and posttranslational processing of multiple forms of tissue-specific exported and intracellular acetylcholinesterase and butyrylcholinesterase. These findings demonstrate that polymorphic cholinesterases are produced from nascent polypeptide products which undergo further posttranslational processing events in a tissue-specific manner before they become mature compartmentalized cholinesterases.
ESTHER : Soreq_1986_Cell.Mol.Neurobiol_6_227
PubMedSearch : Soreq_1986_Cell.Mol.Neurobiol_6_227
PubMedID: 3802131

Title : Synthesis of plasma proteins in fetal, adult, and neoplastic human brain tissue - Dziegielewska_1986_Dev.Biol_115_93
Author(s) : Dziegielewska KM , Saunders NR , Schejter EJ , Zakut H , Zevin-Sonkin D , Zisling R , Soreq H
Ref : Developmental Biology , 115 :93 , 1986
Abstract : The synthesis of plasma proteins directed by mRNA from human brain tissues was studied by combining in vitro or in ovo translation of mRNAs with crossed immunoelectrophoresis of the mRNA-directed labeled polypeptides, followed by autoradiography of the washed plates. Poly(A)-containing mRNA was prepared from different developmental stages of fetal and postnatal human brain and also from primary glioblastomas and meningiomas. Several plasma protein-like polypeptides were identified in the autoradiographs by their migration coordinates in the two-dimensional gels, compared with immunoprecipitates formed by mature, unlabeled, stainable proteins. These included polypeptides migrating like Gc globulin, haptoglobin, fibrinogen, alpha-fetoprotein, transferrin, cholinesterase, and alpha 2-macroglobulin; other, yet unidentified plasma proteins, were also observed. In general, the synthesis of these plasma proteins appeared to be more pronounced in fetal and neoplastic brain tissues than in postnatal tissues. However, clear immunoprecipitates for some of these plasma proteins could also be detected in products directed by mRNA from particular regions of mature, normal brains, indicating that some synthesis of plasma proteins takes place in the human brain even as late as 40 years of age. mRNAs for several proteins were also identified in samples of neoplastic brain. mRNA for transferrin was identified in normal fetal and adult brain but not in either the glioblastomas or meningiomas studied. Microinjected Xenopus oocytes, in which post-translational processing occurs as well, were also used to translate fetal brain mRNA. Several plasma proteins could be detected in the translation products which were induced and stored in the oocytes. These included hemopexin, which could not be detected in the in vitro system. Others, such as cholinesterase, were found to be secreted by the oocytes. These findings indicate that different cell types in the human brain may produce and either store or secrete particular plasma proteins at defined stages in their development.
ESTHER : Dziegielewska_1986_Dev.Biol_115_93
PubMedSearch : Dziegielewska_1986_Dev.Biol_115_93
PubMedID: 2422074

Title : A human acetylcholinesterase gene identified by homology to the Ace region of Drosophila - Soreq_1985_Proc.Natl.Acad.Sci.U.S.A_82_1827
Author(s) : Soreq H , Zevin-Sonkin D , Avni A , Hall LM , Spierer P
Ref : Proceedings of the National Academy of Sciences of the United States of America , 82 :1827 , 1985
Abstract : The Ace locus of the Drosophila genome controls biosynthesis of the neurotransmitter-hydrolyzing enzyme acetylcholinesterase (acetylcholine acetylhydrolase, EC We injected the mRNA species hybridizing with DNA fragments from this region into Xenopus oocytes, in which acetylcholinesterase mRNA is translated into active acetylcholinesterase. A 2.0-kilobase (kb) fragment of DNA from this region selectively hybridizes with Drosophila mRNA capable of inducing the biosynthesis of acetylcholinesterase in oocytes. This Drosophila DNA fragment cross-hybridized with human brain poly(A)+ RNA. We therefore used this DNA fragment as a probe for homologous sequence(s) in a human genomic DNA library and thus selected a 13.5-kb human DNA segment. DNA blot-hybridization revealed that a 2.6-kb fragment of this human DNA segment hybridizes with the Drosophila 2.0-kb DNA fragment. Both Drosophila and human fragments hybridized with a human brain mRNA species of about 7.0-kb that was barely detectable in the acetylcholinesterase-deficient HEp carcinoma. A fraction containing mRNA of similar size, extracted from human brain, induced acetylcholinesterase biosynthesis in oocytes. The human DNA fragment also was used in hybridization-selection experiments. In oocytes, hybrid-selected human brain mRNA induced acetylcholinesterase activity that was completely inhibited by 1,5-bis[4-allyldimethylammonium)phenyl]pentan-3-one dibromide but not by tetraisopropyl pyrophosphamide, a differential response to these inhibitors characteristic of "true" human brain acetylcholinesterase. These findings strongly suggest that both the Drosophila and the human DNA fragments are directly involved in controlling acetylcholinesterase biosynthesis.
ESTHER : Soreq_1985_Proc.Natl.Acad.Sci.U.S.A_82_1827
PubMedSearch : Soreq_1985_Proc.Natl.Acad.Sci.U.S.A_82_1827
PubMedID: 3856864

Title : Expression of acetylcholinesterase gene(s) in the human brain: molecular cloning evidence for cross-homologous sequences - Zevin-Sonkin_1985_J.Physiol_80_221
Author(s) : Zevin-Sonkin D , Avni A , Zisling R , Koch R , Soreq H
Ref : The Journal of Physiology , 80 :221 , 1985
Abstract : The regulation of acetylcholinesterase (AChE) in the human brain has been approached at the level of the genome. A human DNA fragment of the length of 2 600 nucleotides was isolated from a human genomic library. This DNA fragment, designated Huache 1R, bears sequence homology to a DNA fragment from the vicinity of the Drosophila Ace region, that controls AChE biosynthesis (Soreq et al., 1985). Polyadenylated RNA from human brain was hybridized with Huache 1R DNA, eluted and microinjected into Xenopus oocytes in the absence or presence of 35S-methionine. The hybrid-selected RNA induced the biosynthesis of active AChE in the oocytes. Immunoprecipitation of labeled oocyte proteins with monoclonal antibodies against human AChE (Fambrough et al., 1982) resulted in the selective precipitation of an 85 000 Mr induced protein, with a similar size to that of the subunit of human brain AChE. These findings show that the Huache 1R DNA hybridizes with human brain AChEmRNA. The Huache 1R fragment was employed to select a collection of 12 homologous phage-cloned human genomic DNA fragments with different restriction patterns. A cDNA library in pBR322 plasmids was prepared from polyadenylated RNA isolated from embryonic brain. This library was also screened using labeled Huache 1R DNA as a probe. Forty-two out of 37 000 colonies were found positive. Several of these were selected for further analyses. Hybrid-selection experiments using DNA from two of the positive plasmid clones showed that these cDNAs also hybridize with AChEmRNA from human brain. DNA blot hybridization revealed homologies between these cDNA chains and the original Huache 1 fragment.
ESTHER : Zevin-Sonkin_1985_J.Physiol_80_221
PubMedSearch : Zevin-Sonkin_1985_J.Physiol_80_221
PubMedID: 3938815

Title : Expression of cholinesterase gene(s) in human brain tissues: translational evidence for multiple mRNA species - Soreq_1984_EMBO.J_3_1371
Author(s) : Soreq H , Zevin-Sonkin D , Razon N
Ref : EMBO Journal , 3 :1371 , 1984
Abstract : To resolve the origin(s) of the molecular heterogeneity of human nervous system cholinesterases (ChEs), we used Xenopus oocytes, which produce biologically active ChE when microinjected with unfractionated brain mRNA. The RNA was prepared from primary gliomas, meningiomas and embryonic brain, each of which expresses ChE activity with distinct substrate specificities and molecular forms. Sucrose gradient fractionation of DMSO-denatured mRNA from these sources revealed three size classes of ChE-inducing mRNAs, sedimenting at approximately 32S, 20S and 9S. The amounts of these different classes of ChE-inducing mRNAs varied between the three tissue sources examined. To distinguish between ChEs produced in oocytes and having different substrate specificities, their activity was determined in the presence of selective inhibitors. Both 'true' (acetylcholine hydrolase, EC and 'pseudo' (acylcholine acylhydrolase, EC multimeric cholinesterase activities were found in the mRNA-injected oocytes. Moreover, human brain mRNAs inducing 'true' and 'pseudo' ChE activities had different size distribution, indicating that different mRNAs might be translated into various types of ChEs. These findings imply that the heterogeneity of ChEs in the human nervous system is not limited to the post-translational level, but extends to the level of mRNA.
ESTHER : Soreq_1984_EMBO.J_3_1371
PubMedSearch : Soreq_1984_EMBO.J_3_1371
PubMedID: 6745236