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Title | Nucleotide-dependent conformational changes in the N-Ethylmaleimide Sensitive Factor (NSF) and their potential role in SNARE complex disassembly. |
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Journal, issue, pages | J Struct Biol, Vol. 177, Issue 2, Page 335-343, Year 2012 |
Publish date | Jan 5, 2012 |
Authors | Arne Moeller / Chunxia Zhao / Michael G Fried / Elizabeth M Wilson-Kubalek / Bridget Carragher / Sidney W Whiteheart / |
PubMed Abstract | Homohexameric, 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 links | J Struct Biol / PubMed:22245547 / PubMed Central |
Methods | EM (single particle) |
Resolution | 16.0 - 20.0 Å |
Structure data | EMDB-2039: EMDB-2040: EMDB-2041: |
Source |
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