Association of l-Glutamic Acid Decarboxylase to the 70-kDa Heat Shock Protein as a Potential Anchoring Mechanism to Synaptic Vesicles
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Recently we have reported that the membrane-associated form of the gamma-aminobutyric acid-synthesizing enzyme, l-glutamate decarboxylase (MGAD), is regulated by the vesicular proton gradient (Hsu, C. C., Thomas, C., Chen, W., Davis, K. M., Foos, T., Chen, J. L., Wu, E., Floor, E., Schloss, J. V., and Wu, J. Y. (1999) J. Biol. Chem. 274, 24366-24371). In this report, several lines of evidence are presented to indicate that l-glutamate decarboxylase (GAD) can become membrane-associated to synaptic vesicles first through complex formation with the heat shock protein 70 family, specifically heat shock cognate 70 (HSC70), followed by interaction with cysteine string protein (CSP), an integral protein of the synaptic vesicle. The first line of evidence comes from purification of MGAD in which HSC70, as identified from amino acid sequencing, co-purified with GAD. Second, in reconstitution studies, HSC70 was found to form complex with GAD(65) as shown by gel mobility shift in non-denaturing gradient gel electrophoresis. Third, in immunoprecipitation studies, again, HSC70 was co-immunoprecipitated with GAD by a GAD(65)-specific monoclonal antibody. Fourth, HSC70 and CSP were co-purified with GAD by specific anti-GAD immunoaffinity columns. Furthermore, studies here suggest that both GAD(65) and GAD(67) are associated with synaptic vesicles along with HSC70 and CSP. Based on these findings, a model is proposed to link anchorage of MGAD to synaptic vesicles in relation to its role in gamma-aminobutyric acid neurotransmission.
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GAD67 and GAD65 form complexes with vGAT on synaptic vesicles, facilitating coupling from GABA synthesis to vesicular packaging.
“It should be noted that, in addition to mediating cytosolic GABA synthesis, which influences cellular GABA levels, GAD67 and GAD65 form complexes that directly interact with vGAT on synaptic vesicles, thereby facilitating the coupling from GABA synthesis to vesicular packaging (51,52).”
From MeCP2 regulates the timing of critical period plasticity that shapes functional connectivity in primary visual cortex (2015) · cites this paper as [52]
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- Authors
- Hsu, C.-C., Davis, K. M., Jin, H., Foos, T., Floor, E., Chen, W., Tyburski, J. B., Yang, C.-Y., Schloss, J. V., & Wu, J.-Y.
- Affiliations
- University of Kansas (US)
- Baylor College of Medicine (US)
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2000-07
- Volume
- 275
- Issue
- 27
- Pages
- 20822-20828
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1979302622
- Cited by
- 69
- References
- 37
- Citations by year
- 2025: 3 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 1 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 1 · 2016: 2 · 2015: 2 · 2014: 4 · 2013: 1 · 2012: 3
- FWCI
- 1.81
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- glutamic acid decarboxylase, Hsc70, HSP70, synaptic vesicles, cysteine string protein, GABA synthesis, protein complex formation, GAD65, GAD67, GABAergic neurotransmission
- MeSH terms
- Animals, Antibodies, Monoclonal, Brain, Carrier Proteins, Cattle, Enzyme Activation, gamma-Aminobutyric Acid, Glutamate Decarboxylase, Humans, Membrane Proteins, Precipitin Tests, Recombinant Proteins, Synaptic Vesicles, Neurotransmitter Agents, HSP70 Heat-Shock Proteins, HSC70 Heat-Shock Proteins, HSP40 Heat-Shock Proteins
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