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Cited by 197Cellular transport and secretionNeuroscience and Neuropharmacology ResearchAdvanced Fluorescence Microscopy Techniques

Single-molecule FRET–derived model of the synaptotagmin 1–SNARE fusion complex

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Choi, Ucheor B, Strop, Pavel, Vrljic, Marija, Chu, Steven, Brunger, Axel T, Weninger, Keith R·2010·Nature Structural & Molecular Biology
DOI 10.1038/nsmb.1763PMC PMC2922927
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Abstract

Synchronous neurotransmission is triggered when Ca(2+) binds to synaptotagmin 1 (Syt1), a synaptic-vesicle protein that interacts with SNAREs and membranes. We used single-molecule fluorescence resonance energy transfer (FRET) between synaptotagmin's two C2 domains to determine that their conformation consists of multiple states with occasional transitions, consistent with domains in random relative motion. SNARE binding results in narrower intrasynaptotagmin FRET distributions and less frequent transitions between states. We obtained an experimentally determined model of the elusive Syt1-SNARE complex using a multibody docking approach with 34 FRET-derived distances as restraints. The Ca(2+)-binding loops point away from the SNARE complex, so they may interact with the same membrane. The loop arrangement is similar to that of the crystal structure of SNARE-induced Ca(2+)-bound Syt3, suggesting a common mechanism by which the interaction between synaptotagmins and SNAREs aids in Ca(2+)-triggered fusion.

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  • The observed smFRET histograms suggested several possible interfaces between Syt1 and the SNARE complex.

    “The observed smFRET histograms28 suggested several possible interfaces between Syt1 and the SNARE complex.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [28]

    The Discussion (page not provided) says that broad, multi-peaked intermolecular smFRET distributions suggest multiple conformations of the Syt1–SNARE complex. It does not say that the histograms suggest several distinct binding interfaces; the Results instead describe a dominant modeled interface. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • Syt1 interacts with the neuronal SNARE complex.

    “Syt1 also interacts with the neuronal SNARE complex based on immunoprecipitation and pull-downs24–27, single molecule fluorescence resonance energy transfer (smFRET)28, and nuclear magnetic resonance29,30 experiments.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [28]

  • Other dynamic or approximate models of the C2AB-SNARE complex obtained by NMR cannot be readily compared with the previous smFRET studies and the results presented here because of differences in conditions, covalent attachment of lanthanide labels, and lack of atomic resolution.

    “Other dynamic or approximate models of the C2AB-SNARE complex were obtained by nuclear magnetic resonance (NMR)29,30, but cannot be readily compared with the previous smFRET studies28 and the results presented here because of differences in conditions, particular covalent attachment of lanthanide labels30, and lack of atomic resolution.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [28]

  • All three interfaces between Syt1 and the SNARE complex fall within the range of smFRET efficiency histograms obtained previously.

    “All three interfaces between Syt1 and the SNARE complex fall within the range of smFRET efficiency histograms obtained previously28 (Extended Data Fig. 4).”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [28]

  • The top smFRET-derived model and the primary interface observed in the crystal structure primarily differed in the orientation of the C2B domain.

    “In retrospect, the top smFRET-derived model28 and the primary interface observed in the crystal structure primarily differed in the orientation of the C2B domain.”

    From Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis (2015) · cites this paper as [28]

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References 57 · Cited by 197

Research works printed but not linked: 41

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Authors
Choi, U. B., Strop, P., Vrljic, M., Chu, S., Brunger, A. T., & Weninger, K. R.
Affiliations
  • North Carolina State University (US)
  • Howard Hughes Medical Institute (US)
  • Lawrence Berkeley National Laboratory (US)
Corresponding author
Weninger, Keith R
ORCID
Venue
Nature Structural & Molecular Biology
Publisher
Springer Science and Business Media LLC
Published
2010-02-21
Volume
17
Issue
3
Pages
318-324
ISSN
  • 1545-9985 (ISSN-L)
  • 1545-9993
DOI
10.1038/nsmb.1763
PMC
PMC2922927
OpenAlex
W2011940688
Cited by
197
References
57
Citations by year
2026: 2 · 2025: 5 · 2024: 4 · 2023: 5 · 2022: 4 · 2021: 5 · 2020: 3 · 2019: 9 · 2018: 13 · 2017: 13 · 2016: 8 · 2015: 19 · 2014: 23 · 2013: 28 · 2012: 24
FWCI
11.75
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Structural & Molecular Biology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Institute of Mental Health (R01-MH63105, R37MH063105, R01MH063105)
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 › Biophysics › Advanced Fluorescence Microscopy Techniques
Keywords
synaptotagmin-1, SNARE complex, synaptic vesicle fusion, C2 domain, conformational change, membrane fusion mechanism
MeSH terms
Animals, Calcium, Chromatography, Affinity, Chromatography, Gel, Chromatography, Ion Exchange, Protein Binding, Recombinant Fusion Proteins, Fluorescence Resonance Energy Transfer, SNARE Proteins, Synaptotagmin I, Rats

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