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Biochim Biophys Acta. 2012 Jan;1823(1):159-71. doi: 10.1016/j.bbamcr.2011.06.003. Epub 2011 Jun 13.

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

Author information

1
Department of Cellular and Molecular Biochemistry, University of Kentucky Medical Center, Lexington, KY 40536-0509, USA. chz024@ucsd.edu

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.

PMID:
21689688
PMCID:
PMC3983028
DOI:
10.1016/j.bbamcr.2011.06.003
[Indexed for MEDLINE]
Free PMC Article

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