PDBsum entry 3j98

Hydrolase

PDB id: 3j98

Contents

Protein chains

(+ 0 more) 676 a.a.

286 a.a.

61 a.a.

66 a.a.

131 a.a.

PDB id:

3j98

Name:

Hydrolase

Title:

Structure of 20s supercomplex determined by single particle cryoelectron microscopy (state iiia)

Structure:

Vesicle-fusing atpase. Chain: a, b, c, d, e, f. Synonym: n-ethylmaleimide-sensitive fusion protein, nem-sensitive fusion protein, vesicular-fusion protein nsf, n-ethylmaleimide sensitive factor. Engineered: yes. Alpha-soluble nsf attachment protein. Chain: h, i, j, g. Synonym: snap-alpha, n-ethylmaleimide-sensitive factor attachment

Source:

Cricetulus griseus. Chinese hamster. Organism_taxid: 10029. Gene: nsf. Expressed in: escherichia coli. Expression_system_taxid: 562. Rattus norvegicus. Rat. Organism_taxid: 10116.

Authors:

M.Zhao,S.Wu,Y.Cheng,A.T.Brunger

Key ref:

M.Zhao et al. (2015). Mechanistic insights into the recycling machine of the SNARE complex. Nature, 518, 61-67. PubMed id: 25581794 DOI: 10.1038/nature14148

Date:

05-Dec-14 Release date: 28-Jan-15

Protein chains

P18708  (NSF_CRIGR) -  Vesicle-fusing ATPase from Cricetulus griseus

Seq: 744 a.a.

Struc: 676 a.a.

Protein chains

P54921  (SNAA_RAT) -  Alpha-soluble NSF attachment protein from Rattus norvegicus

Seq: 295 a.a.

Struc: 286 a.a.

Protein chain

P63045  (VAMP2_RAT) -  Vesicle-associated membrane protein 2 from Rattus norvegicus

Seq: 116 a.a.

Struc: 61 a.a.

Protein chain

P32851  (STX1A_RAT) -  Syntaxin-1A from Rattus norvegicus

Seq: 288 a.a.

Struc: 66 a.a.

Protein chain

P60881  (SNP25_RAT) -  Synaptosomal-associated protein 25 from Rattus norvegicus

Seq: 206 a.a.

Struc: 131 a.a.*

* PDB and UniProt seqs differ at 6 residue positions (black crosses)

Enzyme reactions

• Enzyme class 2: Chains A, B, C, D, E, F: E.C.3.6.4.6 - vesicle-fusing ATPase.
• Reaction: ATP + H2O = ADP + phosphate + H⁺
• Enzyme class 3: Chains H, I, J, G, K, L, M: E.C.?

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1038/nature14148

Nature 518:61-67 (2015)

PubMed id: 25581794

Mechanistic insights into the recycling machine of the SNARE complex.

M.Zhao, S.Wu, Q.Zhou, S.Vivona, D.J.Cipriano, Y.Cheng, A.T.Brunger.

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.