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Cited by 170Cellular transport and secretionNeuroscience and Neuropharmacology ResearchLipid Membrane Structure and Behavior

Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion

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Diao, Jiajie, Grob, Patricia, Cipriano, Daniel J, Kyoung, Minjoung, Zhang, Yunxiang, Shah, Sachi, Nguyen, Amie, Padolina, Mark +6 more·2012·eLife
DOI 10.7554/elife.00109PMC PMC3514886
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Abstract

The molecular underpinnings of synaptic vesicle fusion for fast neurotransmitter release are still unclear. Here, we used a single vesicle-vesicle system with reconstituted SNARE and synaptotagmin-1 proteoliposomes to decipher the temporal sequence of membrane states upon Ca(2+)-injection at 250-500 μM on a 100-ms timescale. Furthermore, detailed membrane morphologies were imaged with cryo-electron microscopy before and after Ca(2+)-injection. We discovered a heterogeneous network of immediate and delayed fusion pathways. Remarkably, all instances of Ca(2+)-triggered immediate fusion started from a membrane-membrane point-contact and proceeded to complete fusion without discernible hemifusion intermediates. In contrast, pathways that involved a stable hemifusion diaphragm only resulted in fusion after many seconds, if at all. When complexin was included, the Ca(2+)-triggered fusion network shifted towards the immediate pathway, effectively synchronizing fusion, especially at lower Ca(2+)-concentration. Synaptic proteins may have evolved to select this immediate pathway out of a heterogeneous network of possible membrane fusion pathways.DOI:http://dx.doi.org/10.7554/eLife.00109.001.

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Authors
Diao, J., Grob, P., Cipriano, D. J., Kyoung, M., Zhang, Y., Shah, S., Nguyen, A., Padolina, M., Srivastava, A., Vrljic, M., Shah, A., Nogales, E., Chu, S., & Brunger, A. T.
Affiliations
  • Howard Hughes Medical Institute (US)
  • Lawrence Berkeley National Laboratory (US)
  • Departments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USA
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2012-12-13
Volume
1
Pages
e00109-e00109
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.00109
PMC
PMC3514886
OpenAlex
W2158350205
Cited by
170
References
2
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FWCI
7.88
Type
journal article
Language
en
Update notices
correction (2015-10-21)
Open access
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  • eLife (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by)
  • OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Institutes of Health (R37-MH63105)
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 › Lipid Membrane Structure and Behavior
Keywords
synaptic vesicle fusion, SNARE proteins, synaptotagmin-1, complexin, calcium-triggered fusion, hemifusion, proteoliposomes
MeSH terms
Action Potentials, Animals, Biological Transport, Calcium, Membrane Fusion, Nerve Tissue Proteins, Synaptic Transmission, Proteolipids, Recombinant Proteins, Synaptic Vesicles, Time Factors, Gene Expression, Adaptor Proteins, Vesicular Transport, Synaptosomal-Associated Protein 25, Syntaxin 1, Synaptotagmin I, Vesicle-Associated Membrane Protein 2, Rats

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