Accessory proteins stabilize the acceptor complex for synaptobrevin, the 1:1 syntaxin/SNAP-25 complex.
Syntaxin/SNAP-25 interactions precede assembly of the ternary SNARE complex that is essential for neurotransmitter release. This binary complex has been difficult to characterize by bulk methods because of the prevalence of a 2:1 dead-end species. Here, using single-molecule fluorescence, we find the structure of the 1:1 syntaxin/SNAP-25 binary complex is variable, with states changing on the second timescale. One state corresponds to a parallel three-helix bundle, whereas other states show one of the SNAP-25 SNARE domains dissociated. Adding synaptobrevin suppresses the dissociated helix states. Remarkably, upon addition of complexin, Munc13, Munc18, or synaptotagmin, a similar effect is observed. Thus, the 1:1 binary complex is a dynamic acceptor for synaptobrevin binding, and accessory proteins stabilize this acceptor. In the cellular environment the binary complex is actively maintained in a configuration where it can rapidly interact with synaptobrevin, so formation is not likely a limiting step for neurotransmitter release.
Weninger
Keith
K
Department of Physics, North Carolina State University, Raleigh, NC 27695-8202, USA.
Bowen
Mark E
ME
Choi
Ucheor B
UB
Chu
Steven
S
Brunger
Axel T
AT
D031541:Fluorescence Resonance Energy Transfer; D008051:Lipid Bilayers / Q000378:metabolism; D017434:Protein Structure, Tertiary; D050765:Qa-SNARE Proteins / Q000737:chemistry* / Q000378:metabolism; D050683:R-SNARE Proteins / Q000378:metabolism*; D050600:SNARE Proteins / Q000378:metabolism*; D050825:Synaptosomal-Associated Protein 25 / Q000737:chemistry* / Q000378:metabolism
D008051:Lipid Bilayers; D050765:Qa-SNARE Proteins; D050683:R-SNARE Proteins; D050600:SNARE Proteins; D050825:Synaptosomal-Associated Protein 25