Structural characterization of full-length NSF and 20S particles.
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.
State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing, China.
Lei-Fu
LF
Chang
Song
S
Chen
Cui-Cui
CC
Liu
Xijiang
X
Pan
Jiansen
J
Jiang
Xiao-Chen
XC
Bai
Xin
X
Xie
Hong-Wei
HW
Wang
Sen-Fang
SF
Sui
D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D006224:Cricetinae; D003412:Cricetulus; D020285:Cryoelectron Microscopy; D008958:Models, Molecular; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D050600:SNARE Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D040681:Structural Homology, Protein; D013379:Substrate Specificity
D033942:Adaptor Proteins, Vesicular Transport; D050600:SNARE Proteins; D000244:Adenosine Diphosphate; D052066:N-Ethylmaleimide-Sensitive Proteins