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Structure paper

TitleMechanistic insights into the SNARE complex disassembly.
Journal, issue, pagesSci Adv, Vol. 5, Issue 4, Page eaau8164, Year 2019
Publish dateApr 10, 2019
AuthorsXuan Huang / Shan Sun / Xiaojing Wang / Fenghui Fan / Qiang Zhou / Shan Lu / Yong Cao / Qiu-Wen Wang / Meng-Qiu Dong / Jun Yao / Sen-Fang Sui /
PubMed AbstractNSF (-ethylmaleimide-sensitive factor) and α-SNAP (α-soluble NSF attachment protein) bind to the SNARE (soluble NSF attachment protein receptor) complex, the minimum machinery to mediate membrane ...NSF (-ethylmaleimide-sensitive factor) and α-SNAP (α-soluble NSF attachment protein) bind to the SNARE (soluble NSF attachment protein receptor) complex, the minimum machinery to mediate membrane fusion, to form a 20S complex, which disassembles the SNARE complex for reuse. We report the cryo-EM structures of the α-SNAP-SNARE subcomplex and the NSF-D1D2 domain in the 20S complex at 3.9- and 3.7-Å resolutions, respectively. Combined with the biochemical and electrophysiological analyses, we find that α-SNAPs use R116 through electrostatic interactions and L197 through hydrophobic interactions to apply force mainly on two positions of the VAMP protein to execute disassembly process. Furthermore, we define the interaction between the amino terminus of the SNARE helical bundle and the pore loop of the NSF-D1 domain and demonstrate its essential role as a potential anchor for SNARE complex disassembly. Our studies provide a rotation model of α-SNAP-mediated disassembly of the SNARE complex.
External linksSci Adv / PubMed:30989110 / PubMed Central
MethodsEM (single particle)
Resolution3.7 - 8.241905 Å
Structure data

9697

6ip1

EMDB-9697, PDB-6ip1:
alpha-SNAP-SNARE subcomplex in the whole 20S complex
Method: EM (single particle) / Resolution: 3.9 Å

9698

6ip2

EMDB-9698, PDB-6ip2:
NSF-D1D2 part in the whole 20S complex
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-9723:
state1 of the whole 20S complex
Method: EM (single particle) / Resolution: 7.51 Å

EMDB-9724:
state2 of the whole 20S complex
Method: EM (single particle) / Resolution: 8.241905 Å

EMDB-9725:
state3 of the whole 20S complex
Method: EM (single particle) / Resolution: 8.241905 Å

EMDB-9726:
state4 of the whole 20S complex
Method: EM (single particle) / Resolution: 8.045669 Å

EMDB-9727:
state5 of the whole 20S complex
Method: EM (single particle) / Resolution: 7.858561 Å

EMDB-9728:
state6 of the whole 20S complex
Method: EM (single particle) / Resolution: 7.509291 Å

EMDB-9729:
the whole 20S complex at the resolution of 4.64 angstrom
Method: EM (single particle) / Resolution: 4.645476 Å

Chemicals

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

Source
  • rattus norvegicus (Norway rat)
  • bos taurus (cattle)
  • cricetulus griseus (Chinese hamster)
KeywordsMEMBRANE PROTEIN / membrane fusion / ATPase / HYDROLASE

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