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PMID: 25581794 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Mechanistic insights into the recycling machine of the SNARE complex.

Nature ·Vol. 518 ·No. 7537 ·2015-02-05 ·Pages 61-7

Zhao M, Wu S, Zhou Q, Vivona S, Cipriano DJ, Cheng Y, Brunger AT

Abstract

Evolutionarily conserved SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptors) proteins form a complex that drives membrane fusion in eukaryotes. The ATPase NSF (N-ethylmaleimide sensitive factor), together with SNAPs (soluble NSF attachment protein), disassembles the SNARE complex into its protein components, making individual SNAREs available for subsequent rounds of fusion. Here we report structures of ATP- and ADP-bound NSF, and the NSF/SNAP/SNARE (20S) supercomplex determined by single-particle electron cryomicroscopy at near-atomic to sub-nanometre resolution without imposing symmetry. Large, potentially force-generating, conformational differences exist between ATP- and ADP-bound NSF. The 20S supercomplex exhibits broken symmetry, transitioning from six-fold symmetry of the NSF ATPase domains to pseudo four-fold symmetry of the SNARE complex. SNAPs interact with the SNARE complex with an opposite structural twist, suggesting an unwinding mechanism. The interfaces between NSF, SNAPs, and SNAREs exhibit characteristic electrostatic patterns, suggesting how one NSF/SNAP species can act on many different SNARE complexes.

MeSH Terms
Adenosine Diphosphate/metabolism Adenosine Triphosphate/metabolism Animals Cricetulus Cryoelectron Microscopy Models, Molecular Multiprotein Complexes/chemistry,metabolism,ultrastructure N-Ethylmaleimide-Sensitive Proteins/chemistry,metabolism,ultrastructure Protein Binding Protein Structure, Tertiary Rats SNARE Proteins/chemistry,metabolism,ultrastructure Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins/chemistry,metabolism,ultrastructure
Chemicals
Multiprotein Complexes SNARE Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Adenosine Diphosphate Adenosine Triphosphate N-Ethylmaleimide-Sensitive Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Zhao Minglei
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
Wu Shenping
Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA.
Zhou Qiangjun
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
Vivona Sandro
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
Cipriano Daniel J
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
Cheng Yifan
Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA.
Brunger Axel T
1] Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA [2] Department of Neurology and Neurological Sciences, Department of Structural Biology, Department of Photon Science, Stanford University, Stanford, California 94305, USA.
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Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature14148
Published
2015-02-05
Epub
2015-00-12
Pages
61-7
Language
English
Region
England
NLM ID
0410462
PMCID
PMC4320033
Subset
IM
Grants
NIAID NIH HHS · 5-U01AI082051-05 · United States
NIGMS NIH HHS · R01GM082893 · United States
NIAID NIH HHS · U01 AI082051 · United States
NIGMS NIH HHS · P50GM082250 · United States
NIMH NIH HHS · R37 MH063105 · United States
NIMH NIH HHS · R37MH63105 · United States
Howard Hughes Medical Institute · United States
NIGMS NIH HHS · P50 GM082250 · United States
NIGMS NIH HHS · R01GM098672 · United States
NIGMS NIH HHS · R01 GM082893 · United States
NIGMS NIH HHS · R01 GM098672 · United States
Databases
PDB
3J94, 3J95, 3J96, 3J97, 3J98, 3J99
External Links
PubMed source DOI: 10.1038/nature14148 PMC Full Text View journal details
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