1. Microbiology and Infectious Disease
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NSF-mediated disassembly of on and off-pathway SNARE complexes and inhibition by complexin

  1. Ucheor B Choi
  2. Minglei Zhao
  3. K Ian White
  4. Richard A Pfuetzner
  5. Luis Esquivies
  6. Qiangjun Zhou
  7. Axel T Brunger  Is a corresponding author
  1. Stanford University, United States
  2. University of Chicago, United States
Research Article
  • Cited 14
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Cite this article as: eLife 2018;7:e36497 doi: 10.7554/eLife.36497

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 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 sec) and long-lived ({greater than or equal to} 0.32 sec) disassembled states. The long-lived states represent fully disassembled SNARE complex, while the short-lived states correspond to failed disassembly or immediate re-assembly. 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, suggesting a role 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, suggesting a possible regulatory role in disassembly.

Data availability

The EM map associated with this paper has been deposited in the wwPDB under the accession number EMD-8944.

The following data sets were generated

Article and author information

Author details

  1. Ucheor B Choi

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  2. Minglei Zhao

    Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, United States
    Competing interests
    No competing interests declared.
  3. K Ian White

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  4. Richard A Pfuetzner

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  5. Luis Esquivies

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  6. Qiangjun Zhou

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1789-9588
  7. Axel T Brunger

    Department of Molecular and Cellular Physiology, Stanford University, Stanford, United States
    For correspondence
    brunger@stanford.edu
    Competing interests
    Axel T Brunger, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5121-2036

Funding

National Institutes of Health (R37MH63105)

  • Axel T Brunger

Howard Hughes Medical Institute

  • Axel T Brunger

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Vivek Malhotra, The Barcelona Institute of Science and Technology, Spain

Publication history

  1. Received: March 8, 2018
  2. Accepted: July 6, 2018
  3. Accepted Manuscript published: July 9, 2018 (version 1)
  4. Version of Record published: September 10, 2018 (version 2)

Copyright

© 2018, Choi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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