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TitleNucleotide-dependent conformational changes in the N-Ethylmaleimide Sensitive Factor (NSF) and their potential role in SNARE complex disassembly.
Journal, issue, pagesJ Struct Biol, Vol. 177, Issue 2, Page 335-343, Year 2012
Publish dateJan 5, 2012
AuthorsArne Moeller / Chunxia Zhao / Michael G Fried / Elizabeth M Wilson-Kubalek / Bridget Carragher / Sidney W Whiteheart /
PubMed AbstractHomohexameric, N-Ethylmaleimide Sensitive Factor (NSF) disassembles Soluble NSF Attachment Protein Receptor (SNARE) complexes after membrane fusion, an essential step in vesicular trafficking. NSF ...Homohexameric, N-Ethylmaleimide Sensitive Factor (NSF) disassembles Soluble NSF Attachment Protein Receptor (SNARE) complexes after membrane fusion, an essential step in vesicular trafficking. NSF contains three domains (NSF-N, NSF-D1, and NSF-D2), each contributing to activity. We combined electron microscopic (EM) analysis, analytical ultracentrifugation (AU) and functional mutagenesis to visualize NSF's ATPase cycle. 3D density maps show that NSF-D2 remains stable, whereas NSF-N undergoes large conformational changes. NSF-Ns splay out perpendicular to the ADP-bound hexamer and twist upwards upon ATP binding, producing a more compact structure. These conformations were confirmed by hydrodynamic, AU measurements: NSF-ATP sediments faster with a lower frictional ratio (f/f(0)). Hydrodynamic analyses of NSF mutants, with specific functional defects, define the structures underlying these conformational changes. Mapping mutations onto our 3D models allows interpretation of the domain movement and suggests a mechanism for NSF binding to and disassembly of SNARE complexes.
External linksJ Struct Biol / PubMed:22245547 / PubMed Central
MethodsEM (single particle)
Resolution16.0 - 20.0 Å
Structure data

EMDB-2039:
negative stain Electron Microscopy of the N-Ethylmaleimide Sensitive Factor (NSF)
Method: EM (single particle) / Resolution: 20.0 Å

EMDB-2040:
negative stain Electron Microscopy of the N-Ethylmaleimide Sensitive Factor (NSF)
Method: EM (single particle) / Resolution: 17.0 Å

EMDB-2041:
negative stain Electron Microscopy of the N-Ethylmaleimide Sensitive Factor (NSF)
Method: EM (single particle) / Resolution: 16.0 Å

Source
  • Cricetulus griseus (Chinese hamster)

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