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Nat Struct Mol Biol. 2010 Mar;17(3):318-24. doi: 10.1038/nsmb.1763. Epub 2010 Feb 21.

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

Author information

1
Department of Physics, North Carolina State University, Raleigh, North Carolina, USA.

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.

PMID:
20173763
PMCID:
PMC2922927
DOI:
10.1038/nsmb.1763
[Indexed for MEDLINE]
Free PMC Article

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