title: Three αSNAP and 10 ATP molecules are used in SNARE complex disassembly by N-ethylmaleimide-sensitive factor (NSF).
abstract: The fusion of intracellular membranes is driven by the formation of a highly stable four-helix bundle of SNARE proteins embedded in the vesicle and target membranes. N-Ethylmaleimide sensitive factor recycles SNAREs after fusion by binding to the SNARE complex through an adaptor protein, αSNAP, and using the energy of ATP hydrolysis to disassemble the complex. Although only a single molecule of αSNAP binds to a soluble form of the SNARE complex, we find that three molecules of αSNAP are used for SNARE complex disassembly. We describe an engineered αSNAP trimer that supports more efficient SNARE complex disassembly than monomeric αSNAP. Using the trimerized αSNAP, we find that N-ethylmaleimide-sensitive factor hydrolyzes 10 ATP molecules on average to disassemble a single SNARE complex.
authors.affiliation: From the Departments of Structural Biology, Molecular and Cellular Physiology, and.
authors.forename: Niket
authors.initials: N
authors.lastname: Shah
authors.affiliation: From the Departments of Structural Biology, Molecular and Cellular Physiology, and.
authors.forename: Karen N
authors.initials: KN
authors.lastname: Colbert
authors.affiliation: From the Departments of Structural Biology, Molecular and Cellular Physiology, and.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Enos
authors.affiliation: Biochemistry, Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Daniel
authors.initials: D
authors.lastname: Herschlag
authors.affiliation: From the Departments of Structural Biology, Molecular and Cellular Physiology, and bill.weis@stanford.edu.
authors.forename: William I
authors.initials: WI
authors.lastname: Weis
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D016880:Anisotropy; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism; D003412:Cricetulus; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D008561:Membrane Fusion; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000737:chemistry; D011485:Protein Binding; D015202:Protein Engineering; D021381:Protein Transport; D050600:SNARE Proteins / Q000737:chemistry*; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000737:chemistry*
chemical_list: D009711:Nucleotides; D050600:SNARE Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D000255:Adenosine Triphosphate
keywords: ATPase; ATPases Associated with Diverse Cellular Activities (AAA); Intracellular Trafficking; Membrane Trafficking; NSF; Soluble NSF Attachment Protein Receptor (SNARE); alpha-SNAP