Requirements for the catalytic cycle of the N-ethylmaleimide-Sensitive Factor (NSF)

Chunxia Zhao, Everett C. Smith, Sidney W. Whiteheart

Research output: Contribution to journalReview articlepeer-review

49 Scopus citations

Abstract

The N-ethylmaleimide-Sensitive Factor (NSF) was one of the initial members of the ATPases Associated with various cellular Activities Plus (AAA +) family. In this review, we discuss what is known about the mechanism of NSF action and how that relates to the mechanisms of other AAA + proteins. Like other family members, NSF binds to a protein complex (i.e., SNAP-SNARE complex) and utilizes ATP hydrolysis to affect the conformations of that complex. SNAP-SNARE complex disassembly is essential for SNARE recycling and sustained membrane trafficking. NSF is a homo-hexamer; each protomer is composed of an N-terminal domain, NSF-N, and two adjacent AAA-domains, NSF-D1 and NSF-D2. Mutagenesis analysis has established specific roles for many of the structural elements of NSF-D1, the catalytic ATPase domain, and NSF-N, the SNAP-SNARE binding domain. Hydrodynamic analysis of NSF, labeled with (Ni 2+-NTA) 2-Cy3, detected conformational differences in NSF, in which the ATP-bound conformation appears more compact than the ADP-bound form. This indicates that NSF undergoes significant conformational changes as it progresses through its ATP-hydrolysis cycle. Incorporating these data, we propose a sequential mechanism by which NSF uses NSF-N and NSF-D1 to disassemble SNAP-SNARE complexes. We also illustrate how analytical centrifugation might be used to study other AAA + proteins. This article is part of a Special Issue entitled: AAA ATPases: structure and function.

Original languageEnglish
Pages (from-to)159-171
Number of pages13
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1823
Issue number1
DOIs
StatePublished - Jan 2012

Keywords

  • ATPase
  • Membrane trafficking
  • NSF
  • SNAP
  • SNARE

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Requirements for the catalytic cycle of the N-ethylmaleimide-Sensitive Factor (NSF)'. Together they form a unique fingerprint.

Cite this