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TitleStructural principles of SNARE complex recognition by the AAA+ protein NSF.
Journal, issue, pagesElife, Vol. 7, Year 2018
Publish dateSep 10, 2018
AuthorsK Ian White / Minglei Zhao / Ucheor B Choi / Richard A Pfuetzner / Axel T Brunger /
PubMed AbstractThe recycling of SNARE proteins following complex formation and membrane fusion is an essential process in eukaryotic trafficking. A highly conserved AAA+ protein, NSF (-ethylmaleimide sensitive ...The recycling of SNARE proteins following complex formation and membrane fusion is an essential process in eukaryotic trafficking. A highly conserved AAA+ protein, NSF (-ethylmaleimide sensitive factor) and an adaptor protein, SNAP (soluble NSF attachment protein), disassemble the SNARE complex. We report electron-cryomicroscopy structures of the complex of NSF, αSNAP, and the full-length soluble neuronal SNARE complex (composed of syntaxin-1A, synaptobrevin-2, SNAP-25A) in the presence of ATP under non-hydrolyzing conditions at ~3.9 Å resolution. These structures reveal electrostatic interactions by which two αSNAP molecules interface with a specific surface of the SNARE complex. This interaction positions the SNAREs such that the 15 N-terminal residues of SNAP-25A are loaded into the D1 ring pore of NSF via a spiral pattern of interactions between a conserved tyrosine NSF residue and SNAP-25A backbone atoms. This loading process likely precedes ATP hydrolysis. Subsequent ATP hydrolysis then drives complete disassembly.
External linksElife / PubMed:30198481 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 4.4 Å
Structure data

EMDB-9100, PDB-6mdm:
The 20S supercomplex engaging the SNAP-25 N-terminus (class 1)
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-9101, PDB-6mdn:
The 20S supercomplex engaging the SNAP-25 N-terminus (class 2)
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-9102, PDB-6mdo:
The D1 and D2 domain rings of NSF engaging the SNAP-25 N-terminus within the 20S supercomplex (focused refinement on D1/D2 rings, class 1)
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-9103, PDB-6mdp:
The D1 and D2 domain rings of NSF engaging the SNAP-25 N-terminus within the 20S supercomplex (focused refinement on D1/D2 rings, class 2)
Method: EM (single particle) / Resolution: 3.8 Å

Chemicals

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM / Adenosine triphosphate

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

Source
  • cricetulus griseus (Chinese hamster)
  • rattus norvegicus (Norway rat)
KeywordsHYDROLASE / SNARE / NSF / SNAP / ATPase / AAA / disassembly / synapse / membrane fusion / exocytosis

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