Journal: Elife / Year: 2018 Title: NSF-mediated disassembly of on- and off-pathway SNARE complexes and inhibition by complexin. Authors: Ucheor B Choi / Minglei Zhao / K Ian White / Richard A Pfuetzner / Luis Esquivies / Qiangjun Zhou / Axel T Brunger / Abstract: 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 ...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.
History
Deposition
Jul 6, 2018
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Header (metadata) release
Jul 25, 2018
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Map release
Jul 25, 2018
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Update
Dec 11, 2019
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Current status
Dec 11, 2019
Processing site: RCSB / Status: Released
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