Disassembly of All SNARE Complexes by N-Ethylmaleimide-sensitive Factor (NSF) Is Initiated by a Conserved 1:1 Interaction between alpha-Soluble NSF Attachment Protein (SNAP) and SNARE Complex

作者：Vivona Sandro; Cipriano Daniel J; O'Leary Sean; Li Ye Henry; Fenn Timothy D; Brunger Axel T^*

来源：JOURNAL OF BIOLOGICAL CHEMISTRY, 2013, 288(34): 24984-24991.

DOI：10.1074/jbc.M113.489807

Vesicle trafficking in eukaryotic cells is facilitated by SNARE-mediated membrane fusion. The ATPase NSF (N-ethylmaleimide- sensitive factor) and the adaptor protein alpha-SNAP (soluble NSF attachment protein) disassemble all SNARE complexes formed throughout different pathways, but the effect of SNARE sequence and domain variation on the poorly understood disassembly mechanism is unknown. By measuring SNARE-stimulated ATP hydrolysis rates, Michaelis-Menten constants for disassembly, and SNAP-SNARE binding constants for four different ternary SNARE complexes and one binary complex, we found a conserved mechanism, not influenced by N-terminal SNARE domains. alpha-SNAP and the ternary SNARE complex form a 1:1 complex as revealed by multiangle light scattering. We propose a model of NSF-mediated disassembly in which the reaction is initiated by a 1:1 interaction between alpha-SNAP and the ternary SNARE complex, followed by NSF binding. Subsequent additional alpha-SNAP binding events may occur as part of a processive disassembly mechanism.

• 出版日期2013-8-23