Single-molecule FRET–derived model of the synaptotagmin 1–SNARE fusion complex
| Autor: | Axel T. Brunger, Pavel Strop, Marija Vrljic, Ucheor B. Choi, Steven Chu, Keith Weninger |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
endocrine system
Vesicle fusion Recombinant Fusion Proteins membrane fusion protein-protein interactions Chromatography Affinity Article Synaptotagmin 1 synaptic vesicle Synaptotagmins 03 medical and health sciences 0302 clinical medicine Structural Biology Fluorescence Resonance Energy Transfer single molecule FRET Animals Molecular Biology 030304 developmental biology 0303 health sciences SNARE binding Chemistry Synaptotagmin I SNAP25 Single-molecule FRET Chromatography Ion Exchange Rats Crystallography nervous system neurotransmitter release Chromatography Gel Calcium SNARE Proteins SNARE complex 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Nature structural & molecular biology |
| ISSN: | 1545-9985 1545-9993 |
| DOI: | 10.1038/nsmb.1763 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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