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Structure paper

TitleStructural remodeling of target-SNARE protein complexes by NSF enables synaptic transmission.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 8371, Year 2025
Publish dateSep 24, 2025
AuthorsK Ian White / Yousuf A Khan / Kangqiang Qiu / Ashwin Balaji / Sergio Couoh-Cardel / Luis Esquivies / Richard A Pfuetzner / Jiajie Diao / Axel T Brunger /
PubMed AbstractSynaptic vesicles containing neurotransmitters fuse with the plasma membrane upon the arrival of an action potential at the active zone. Multiple proteins organize trans-SNARE complex assembly and ...Synaptic vesicles containing neurotransmitters fuse with the plasma membrane upon the arrival of an action potential at the active zone. Multiple proteins organize trans-SNARE complex assembly and priming, leading to fusion. One target membrane SNARE, syntaxin, forms nanodomains at the active zone, and another, SNAP-25, enters non-fusogenic complexes with it. Here, we reveal mechanistic details of AAA+ protein NSF (N-ethylmaleimide sensitive factor) and SNAP (soluble NSF attachment protein) action before fusion. We show that syntaxin clusters are conserved, that NSF colocalizes with them, and characterize SNARE populations that may exist within or near them using cryo-EM. Supercomplexes of NSF, α-SNAP, and either a syntaxin tetramer or one of two binary complexes of syntaxin-SNAP-25 reveal atomic details of SNARE processing and show how sequential ATP hydrolysis drives disassembly. These results suggest a functional role for syntaxin clusters as reservoirs and a corresponding role for NSF in syntaxin liberation and SNARE protein quality control preceding fusion.
External linksNat Commun / PubMed:40993127 / PubMed Central
MethodsEM (single particle)
Resolution2.95 - 4.5 Å
Structure data

70547

9ojr

EMDB-70547, PDB-9ojr:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 3
Method: EM (single particle) / Resolution: 2.95 Å

70548

9oju

EMDB-70548, PDB-9oju:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 4
Method: EM (single particle) / Resolution: 2.97 Å

70550

9ojz

EMDB-70550, PDB-9ojz:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 5
Method: EM (single particle) / Resolution: 3.39 Å

70553

9ok3

EMDB-70553, PDB-9ok3:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), 3:2:1 alphaSNAP-syntaxin-1a-SNAP-25 subcomplex local refinement, non-hydrolyzing, class 13
Method: EM (single particle) / Resolution: 3.74 Å

70554

9ok5

EMDB-70554, PDB-9ok5:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 16
Method: EM (single particle) / Resolution: 3.29 Å

70559

9okc

EMDB-70559, PDB-9okc:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 17
Method: EM (single particle) / Resolution: 3.67 Å

70594

9olj

EMDB-70594, PDB-9olj:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 18
Method: EM (single particle) / Resolution: 3.52 Å

70598

9olo

EMDB-70598, PDB-9olo:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 19
Method: EM (single particle) / Resolution: 3.56 Å

70608

9om6

EMDB-70608, PDB-9om6:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), 4:2:2 alphaSNAP-syntaxin-1a-SNAP-25 subcomplex local refinement, hydrolyzing, class 23
Method: EM (single particle) / Resolution: 4.14 Å

70616

9omq

EMDB-70616, PDB-9omq:
NSF, substrate free, hydrolyzing, class 24
Method: EM (single particle) / Resolution: 3.86 Å

71437

9paf

EMDB-71437, PDB-9paf:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 6
Method: EM (single particle) / Resolution: 3.82 Å

71438

9pag

EMDB-71438, PDB-9pag:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 7
Method: EM (single particle) / Resolution: 3.62 Å

71458

9pb9

EMDB-71458, PDB-9pb9:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 8
Method: EM (single particle) / Resolution: 3.45 Å

71475

9pba

EMDB-71475, PDB-9pba:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 9
Method: EM (single particle) / Resolution: 3.47 Å

71478

9pbf

EMDB-71478, PDB-9pbf:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 10
Method: EM (single particle) / Resolution: 4.01 Å

71491

9pbv

EMDB-71491, PDB-9pbv:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 11
Method: EM (single particle) / Resolution: 3.91 Å

71496

9pc3

EMDB-71496, PDB-9pc3:
21bin20S complex (NSF-alphaSNAP-2:1 syntaxin-1a:SNAP-25), non-hydrolyzing, class 12
Method: EM (single particle) / Resolution: 3.69 Å

71521

9pcx

EMDB-71521, PDB-9pcx:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 14
Method: EM (single particle) / Resolution: 4.03 Å

71522

9pcz

EMDB-71522, PDB-9pcz:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 15
Method: EM (single particle) / Resolution: 3.65 Å

71523

9pd1

EMDB-71523, PDB-9pd1:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 20
Method: EM (single particle) / Resolution: 4.5 Å

71529

9pd8

EMDB-71529, PDB-9pd8:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 21
Method: EM (single particle) / Resolution: 4.23 Å

71530

9pdb

EMDB-71530, PDB-9pdb:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 22
Method: EM (single particle) / Resolution: 3.83 Å

71533

9pdd

EMDB-71533, PDB-9pdd:
22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a:SNAP-25), hydrolyzing, class 29
Method: EM (single particle) / Resolution: 4.16 Å

71591

9pf2

EMDB-71591, PDB-9pf2:
NSF, substrate free, hydrolyzing, class 25
Method: EM (single particle) / Resolution: 3.88 Å

71598

9pfc

EMDB-71598, PDB-9pfc:
NSF, substrate free, hydrolyzing, class 26
Method: EM (single particle) / Resolution: 3.15 Å

71600

9pff

EMDB-71600, PDB-9pff:
Min22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a H3:SNAP-25 SN1), non-hydrolyzing, class 27
Method: EM (single particle) / Resolution: 3.09 Å

71601

9pfg

EMDB-71601, PDB-9pfg:
Min22bin20S complex (NSF-alphaSNAP-2:2 syntaxin-1a H3:SNAP-25 SN1), 4:2:2 alphaSNAP-syntaxin-1a H3-SNAP-25 SN1 subcomplex local refinement, non-hydrolyzing, class 28
Method: EM (single particle) / Resolution: 3.58 Å

Chemicals

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

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

ChemComp-HOH:
WATER

ChemComp-MG:
Unknown entry

ChemComp-PO4:
PHOSPHATE ION

Source
  • rattus norvegicus (Norway rat)
  • cricetulus griseus (Chinese hamster)
  • rattus rattus (black rat)
KeywordsHYDROLASE / ATPase / SNARE / hydrolysis / disassembly / translocation / exocytosis / neurotransmitter release / synapse / synaptic transmission / membrane fusion

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