Structural characterization of full-length NSF and 20S particles

Nat Struct Mol Biol. 2012 Feb 5;19(3):268-75. doi: 10.1038/nsmb.2237.


The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Vesicular Transport / chemistry*
  • Adaptor Proteins, Vesicular Transport / metabolism
  • Adaptor Proteins, Vesicular Transport / ultrastructure
  • Adenosine Diphosphate / chemistry
  • Adenosine Diphosphate / metabolism
  • Animals
  • Cricetinae
  • Cricetulus
  • Cryoelectron Microscopy
  • Models, Molecular
  • N-Ethylmaleimide-Sensitive Proteins / chemistry*
  • N-Ethylmaleimide-Sensitive Proteins / metabolism
  • N-Ethylmaleimide-Sensitive Proteins / ultrastructure
  • Protein Binding
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary
  • SNARE Proteins / chemistry*
  • SNARE Proteins / metabolism
  • SNARE Proteins / ultrastructure
  • Structural Homology, Protein
  • Substrate Specificity


  • Adaptor Proteins, Vesicular Transport
  • SNARE Proteins
  • Adenosine Diphosphate
  • N-Ethylmaleimide-Sensitive Proteins