Nucleotide-dependent conformational changes in the N-Ethylmaleimide Sensitive Factor (NSF) and their potential role in SNARE complex disassembly

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Arne Möller
  • Chunxia Zhao, Denmark
  • Michael G Fried, Denmark
  • Elizabeth M Wilson-Kubalek, Denmark
  • Bridget Carragher
  • ,
  • Sidney W Whiteheart, Denmark

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.

Original languageEnglish
JournalJournal of Structural Biology
Pages (from-to)335-43
Number of pages9
Publication statusPublished - Feb 2012
Externally publishedYes

    Research areas

  • Adenine Nucleotides, Amino Acid Substitution, Animals, CHO Cells, Cricetinae, Microscopy, Electron, Models, Molecular, N-Ethylmaleimide-Sensitive Proteins, Protein Binding, Protein Structure, Quaternary, Protein Structure, Tertiary, SNARE Proteins, Surface Properties, Ultracentrifugation

See relations at Aarhus University Citationformats

ID: 77176210