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Cited by 98Cellular transport and secretionNeuroscience and Neuropharmacology ResearchExtracellular vesicles in disease

Reconstituted synaptotagmin I mediates vesicle docking, priming, and fusion

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Wang, Zhao, Liu, Huisheng, Gu, Yiwen, Chapman, Edwin R.·2011·Journal of Cell Biology
DOI 10.1083/jcb.201104079PMC PMC3246889
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Abstract

The synaptic vesicle protein synaptotagmin I (syt) promotes exocytosis via its ability to penetrate membranes in response to binding Ca2+ and through direct interactions with SNARE proteins. However, studies using full-length (FL) membrane-embedded syt in reconstituted fusion assays have yielded conflicting results, including a lack of effect, or even inhibition of fusion, by Ca2+. In this paper, we show that reconstituted FL syt promoted rapid docking of vesicles (<1 min) followed by a priming step (3–9 min) that was required for subsequent Ca2+-triggered fusion between v- and t-SNARE liposomes. Moreover, fusion occurred only when phosphatidylinositol 4,5-bisphosphate was included in the target membrane. This system also recapitulates some of the effects of syt mutations that alter synaptic transmission in neurons. Finally, we demonstrate that the cytoplasmic domain of syt exhibited mixed agonist/antagonist activity during regulated membrane fusion in vitro and in cells. Together, these findings reveal further convergence of reconstituted and cell-based systems.

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  • Subsequent work revealed major differences between using the soluble C2AB fragment and reconstituted, full-length Syt1.

    “Moreover, subsequent work revealed major differences between using the soluble C2AB fragment and reconstituted, full-length Syt184,85.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [85]

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Authors
Wang, Z., Liu, H., Gu, Y., & Chapman, E. R.
Affiliations
Howard Hughes Medical Institute (US)
Corresponding author
Chapman, Edwin R.
ORCID
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2011-12-19
Volume
195
Issue
7
Pages
1159-1170
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.201104079
PMC
PMC3246889
OpenAlex
W2128292852
Cited by
98
References
2
Citations by year
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FWCI
5.37
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Institute of Mental Health (MH61876, R01MH061876)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
Keywords
synaptotagmin-1, synaptic vesicle docking, exocytosis, membrane fusion, SNARE proteins, calcium-triggered fusion, phosphatidylinositol 4,5-bisphosphate, vesicle priming, cytoplasmic domain, regulated exocytosis, synaptic transmission
MeSH terms
Animals, Calcium, Cytoplasm, Membrane Fusion, Mutation, Neurons, Phosphatidylserines, Synaptic Vesicles, Time Factors, Mice, Knockout, Phosphatidylinositol 4,5-Diphosphate, SNARE Proteins, Synaptotagmin I, Mice

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