title: Disassembly of all SNARE complexes by N-ethylmaleimide-sensitive factor (NSF) is initiated by a conserved 1:1 interaction between α-soluble NSF attachment protein (SNAP) and SNARE complex.
abstract: Vesicle trafficking in eukaryotic cells is facilitated by SNARE-mediated membrane fusion. The ATPase NSF (N-ethylmaleimide-sensitive factor) and the adaptor protein α-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. α-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 α-SNAP and the ternary SNARE complex, followed by NSF binding. Subsequent additional α-SNAP binding events may occur as part of a processive disassembly mechanism.
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University Medical School, Stanford, California 94305, USA.
authors.forename: Sandro
authors.initials: S
authors.lastname: Vivona
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Cipriano
authors.forename: Seán
authors.initials: S
authors.lastname: O'Leary
authors.forename: Ye Henry
authors.initials: YH
authors.lastname: Li
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Fenn
authors.forename: Axel T
authors.initials: AT
authors.lastname: Brunger
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D006224:Cricetinae; D003412:Cricetulus; D008956:Models, Chemical; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D050600:SNARE Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D046912:Multiprotein Complexes; D050600:SNARE Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D000255:Adenosine Triphosphate; D052066:N-Ethylmaleimide-Sensitive Proteins; C498737:Nsf protein, rat
keywords: ATPases; Biophysics; Biosensors; Circular Dichroism (CD); Enzyme Kinetics; Fluorescence; Membrane Fusion; Protein-Protein Interactions; SNARE Proteins; Structural Biology