NSF-mediated disassembly of on- and off-pathway SNARE complexes and inhibition by complexin.

Autor:	Choi UB; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States., Zhao M; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, United States., White KI; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States., Pfuetzner RA; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States., Esquivies L; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States., Zhou Q; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States., Brunger AT; Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States.; Department of Neurology and Neurological Sciences, Stanford University, Stanford, United States.; Department of Structural Biology, Stanford University, Stanford, United States.; Department of Photon Science, Stanford University, Stanford, United States.; Howard Hughes Medical Institute, Stanford University, Stanford, United States.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2018 Jul 09; Vol. 7. Date of Electronic Publication: 2018 Jul 09.
DOI:	10.7554/eLife.36497
Abstrakt:	SNARE complex disassembly by the ATPase NSF is essential for neurotransmitter release and other membrane trafficking processes. We developed a single-molecule FRET assay to monitor repeated rounds of NSF-mediated disassembly and reassembly of individual SNARE complexes. For ternary neuronal SNARE complexes, disassembly proceeds in a single step within 100 msec. We observed short- (<0.32 s) and long-lived (≥0.32 s) disassembled states. The long-lived states represent fully disassembled SNARE complex, while the short-lived states correspond to failed disassembly or immediate reassembly. Either high ionic strength or decreased αSNAP concentration reduces the disassembly rate while increasing the frequency of short-lived states. NSF is also capable of disassembling anti-parallel ternary SNARE complexes, implicating it in quality control. Finally, complexin-1 competes with αSNAP binding to the SNARE complex; addition of complexin-1 has an effect similar to that of decreasing the αSNAP concentration, possibly differentially regulating cis and trans SNARE complexes disassembly. Competing Interests: UC, MZ, KW, RP, LE, QZ No competing interests declared, AB Reviewing editor, eLife (© 2018, Choi et al.)
Databáze:	MEDLINE
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