Affinage

NSF

Vesicle-fusing ATPase · UniProt P46459

Length
744 aa
Mass
82.6 kDa
Annotated
2026-04-29
100 papers in source corpus 51 papers cited in narrative 51 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NSF is a homo-hexameric AAA+ ATPase that functions as a universal SNARE complex disassembly machine, recycling SNARE proteins after membrane fusion to enable successive rounds of vesicle trafficking across the secretory, endosomal, and lysosomal pathways (PMID:2071670, PMID:25814585, PMID:14617820). Together with its adaptor proteins alpha- and beta-SNAP, NSF binds cis-SNARE complexes to form a 20S supercomplex in which SNAP-25 N-terminal residues are threaded into the NSF D1 ring pore; ATP hydrolysis builds internal tension that is released in a spring-loaded burst (~20 ms), disassembling the SNARE bundle in a single mechanochemical step (PMID:25814585, PMID:30198481, PMID:29985126). Trans-SNARE complexes committed to fusion are functionally resistant to NSF/SNAP-mediated disassembly, and accessory factors including Munc18-1, Munc13-1, complexin-1, and synaptotagmin-1 further protect trans-SNAREs from de-priming (PMID:10831610, PMID:30657450). Beyond canonical SNARE recycling, NSF directly interacts with the GluR2 AMPA receptor subunit to maintain surface receptor expression and regulate synaptic plasticity, and can disassemble non-SNARE protein complexes such as PICK1–GluR2 via the same ATP-hydrolysis-dependent mechanism (PMID:9697854, PMID:11931741, PMID:10571232).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1988 High

    The identification of SEC18 as an essential cytoplasmic ATPase required for ER-to-Golgi transport established that a soluble, NEM-sensitive factor is a core component of the vesicular trafficking machinery.

    Evidence Gene cloning by complementation and cell fractionation of yeast sec18 mutants

    PMID:3054509

    Open questions at the time
    • Oligomeric state unknown
    • Catalytic mechanism uncharacterized
    • Substrate identity not determined
  2. 1990 High

    Purification of alpha-, beta-, and gamma-SNAP as obligate adaptors that recruit NSF to membranes, and identification of membrane-bound SNAP receptors (SNAREs), revealed the tripartite NSF–SNAP–SNARE machinery for membrane fusion.

    Evidence Biochemical purification with in vitro Golgi transport assay and yeast SEC17 complementation

    PMID:2111733 PMID:8455721

    Open questions at the time
    • Structural basis of SNAP–SNARE recognition unknown
    • How NSF uses ATP to act on SNAREs not determined
  3. 1994 High

    Kinetic analysis of NSF ATPase activity showed two distinct ATPase sites (D1 and D2), with SNAP binding activating the low-affinity site by dramatically lowering its Km, explaining how the adaptor-substrate complex switches on NSF catalysis.

    Evidence In vitro ATPase assays with recombinant NSF and SNAP proteins

    PMID:7961908 PMID:9362506

    Open questions at the time
    • Which domain (D1 vs D2) drives disassembly not resolved
    • Single-turnover mechanism unknown
  4. 1997 High

    Electron microscopy and biophysical analyses established NSF as a hexameric cylinder whose conformation is nucleotide-dependent, and visualized the asymmetric 20S complex with alpha-SNAP bridging NSF to the parallel SNARE bundle, providing the first structural framework for the disassembly machine.

    Evidence Quick-freeze/deep-etch EM with epitope labels, analytical ultracentrifugation, MALS

    PMID:9267032 PMID:9624162

    Open questions at the time
    • Atomic-resolution structure lacking
    • Mechanism of force transmission to SNARE complex unknown
  5. 1997 High

    Reconstituted yeast vacuole fusion demonstrated that Sec18p/NSF acts at an early 'priming' step before Rab-dependent docking, ordering NSF function upstream of the actual fusion event and establishing its role as a recycling factor rather than a fusogen.

    Evidence In vitro yeast vacuole fusion with order-of-addition epistasis using antibodies and recombinant proteins

    PMID:8670830 PMID:9015301 PMID:9015302

    Open questions at the time
    • Molecular identity of primed state unclear
    • Whether priming equals SNARE disassembly not directly shown
  6. 1998 High

    Discovery that NSF directly binds the GluR2 AMPA receptor C-terminal domain independently of SNAREs, and that blocking this interaction reduces AMPAR-mediated synaptic transmission, revealed a non-SNARE chaperone function of NSF in neurons.

    Evidence Direct binding assays with mutagenesis, intracellular peptide and antibody loading with electrophysiology in hippocampal neurons

    PMID:10399941 PMID:9697854 PMID:9697855

    Open questions at the time
    • Whether NSF uses the same ATPase cycle on GluR2 as on SNAREs not clear
    • Structural basis of NSF–GluR2 interaction not resolved
  7. 2000 High

    Reconstituted liposome fusion showed that trans-SNARE complexes are resistant to NSF/SNAP disassembly once formed, explaining how productive fusion intermediates survive in a cellular environment favoring SNARE disassembly.

    Evidence In vitro liposome fusion assay with NSF/alpha-SNAP addition at different stages

    PMID:10831610

    Open questions at the time
    • Structural basis of trans-SNARE resistance not determined
    • Whether resistance is absolute or kinetic not resolved
  8. 2001 High

    Genetic analysis of Drosophila comatose NSF mutants proved that NSF acts after fusion to recycle SNARE complexes between exocytosis and endocytosis, definitively placing NSF downstream of the fusion event in vivo.

    Evidence Drosophila temperature-sensitive NSF alleles, SNARE complex immunoblotting, double mutant epistasis with shibire

    PMID:11593041 PMID:9852561

    Open questions at the time
    • Whether NSF has any pre-fusion role in vivo remains debated
    • Temporal resolution of SNARE recycling kinetics in vivo lacking
  9. 2002 High

    NSF was shown to disassemble non-SNARE complexes (PICK1–GluR2) via the same ATP-hydrolysis-dependent mechanism used for SNAREs, broadening its functional scope to a general protein complex disassembly chaperone in neurons.

    Evidence Co-immunoprecipitation and in vitro disassembly assay comparing ATP vs ATPγS conditions, neuronal overexpression

    PMID:11931741

    Open questions at the time
    • Full repertoire of non-SNARE substrates unknown
    • Selectivity mechanism for non-SNARE targets not characterized
  10. 2002 High

    Dissection of NSF's role at GluR2 binding in relation to AP2 and LTD showed that NSF maintains basal AMPAR surface pools while AP2 clathrin adaptor mediates NMDA-triggered internalization, separating NSF-dependent surface stabilization from activity-dependent endocytosis.

    Evidence GluR2 point mutants selectively disrupting NSF vs AP2 binding, hippocampal slice LTD electrophysiology

    PMID:10571232 PMID:12011465 PMID:12441055

    Open questions at the time
    • How NSF prevents constitutive GluR2 endocytosis mechanistically is unclear
    • Whether NSF–GluR2 interaction is SNAP-dependent in vivo not fully resolved
  11. 2015 High

    Single-molecule measurements revealed that NSF disassembles a SNARE complex in one ATP-turnover cycle via a spring-loaded mechanism: tension accumulates over seconds following phosphate release and is discharged in a ~20 ms burst, providing the first direct observation of the NSF mechanochemical cycle.

    Evidence Single-molecule fluorescence spectroscopy and magnetic tweezers

    PMID:25814585

    Open questions at the time
    • How many ATPs per protomer are hydrolyzed per cycle not settled
    • Whether the spring-loaded mechanism applies to non-SNARE substrates untested
  12. 2018 High

    Near-atomic cryo-EM of the 20S supercomplex revealed that two alpha-SNAP molecules bridge NSF to the SNARE bundle via electrostatic contacts, and SNAP-25 N-terminal residues are threaded into the NSF D1 pore in a spiral arrangement, providing the structural basis for substrate engagement prior to hydrolysis.

    Evidence Cryo-EM at ~3.9 Å resolution of NSF/2×alpha-SNAP/neuronal SNARE complex

    PMID:30198481

    Open questions at the time
    • Post-hydrolysis conformational trajectory not captured
    • Structures with different SNARE complexes lacking
  13. 2019 High

    Reconstitution with the full complement of synaptic release factors showed that Munc18-1, Munc13-1, complexin-1, and synaptotagmin-1 all protect trans-SNARE complexes from NSF/SNAP-mediated de-priming, explaining how the cell balances recycling against premature disassembly.

    Evidence In vitro trans-SNARE complex formation assay with purified NSF-alpha-SNAP and synaptic regulators

    PMID:30657450

    Open questions at the time
    • Quantitative contributions of each protective factor in vivo unknown
    • Whether the same protection applies at non-synaptic trafficking steps untested

Open questions

Synthesis pass · forward-looking unresolved questions
  • The full post-hydrolysis conformational trajectory of NSF during the spring-loaded burst, the structural basis for substrate selectivity among different SNARE and non-SNARE complexes, and the in vivo regulation of NSF activity across diverse trafficking pathways remain unresolved.
  • No time-resolved structural snapshots of the power stroke
  • Structural basis for NSF–GluR2 interaction not determined
  • In vivo stoichiometry and regulation of NSF at distinct trafficking stations unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 4 GO:0140657 ATP-dependent activity 4 GO:0044183 protein folding chaperone 2
Localization
GO:0005794 Golgi apparatus 4 GO:0005829 cytosol 2 GO:0031410 cytoplasmic vesicle 2
Pathway
R-HSA-5653656 Vesicle-mediated transport 7 R-HSA-112316 Neuronal System 4 R-HSA-1852241 Organelle biogenesis and maintenance 3 R-HSA-392499 Metabolism of proteins 3
Partners
ARRB1GRIA2LRRK2NAPANAPBNAPGPICK1STX5
Complex memberships
20S complex (NSF hexamer / alpha-SNAP / SNARE bundle)NSF homohexamer

Evidence

Reading pass · 51 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1990 Alpha-SNAP, beta-SNAP, and gamma-SNAP were purified as NSF attachment proteins required for NSF binding to Golgi membranes; SNAP activity is required during the membrane fusion stage of intra-Golgi transport and is conserved in yeast (SEC17). Biochemical purification, in vitro transport assay, yeast complementation Cell High 2111733
1988 SEC18 (yeast NSF ortholog) is an essential, hydrophilic cytoplasmic protein of ~84 kDa required for secretory protein transport between the ER and Golgi complex; it associates with a 100,000×g pellet fraction consistent with transient vesicle binding. Gene cloning by complementation, gene disruption, cell fractionation, pulse-chase Molecular and cellular biology High 3054509
1991 Sec18p/NSF function is required sequentially for protein transport from the ER to the Golgi, through multiple Golgi compartments, and from the Golgi to the cell surface, defining at least three functionally distinct Golgi compartments in yeast. Temperature-sensitive yeast mutants, pulse-chase protein transport assays The Journal of cell biology High 2071670
1993 Alpha- and gamma-SNAP act synergistically in intra-Golgi transport; beta-SNAP is a brain-specific isoform of alpha-SNAP; SNAPs enable NSF to bind to target membranes and specificity is conferred by SNAP receptors (SNAREs) on the membranes being fused. cDNA cloning, in vitro transport assay, tissue expression analysis Nature High 8455721
1997 NSF is a hollow 10×16 nm cylindrical hexamer; its conformation depends on nucleotide binding, converting to a 'splayed' protease-sensitive form when nucleotide-depleted. The ternary SNARE complex (syntaxin/SNAP-25/synaptobrevin) is a rod with syntaxin and synaptobrevin aligned in parallel with membrane anchors at the same end; alpha-SNAP and NSF form an asymmetric '20S' complex by binding one end of the SNARE rod. Quick-freeze/deep-etch electron microscopy with epitope tags, antibodies, and maltose-binding protein markers Cell High 9267032
1994 NSF displays complex ATPase kinetics consistent with two ATPase domains; SNAPs (adsorbed to surface) enhance NSF ATPase activity primarily by decreasing the Km of the low-affinity ATPase site ~100-fold, acting as a molecular switch to activate the normally dormant site. In vitro ATPase assay with recombinant His6-NSF and His6-SNAP proteins The Journal of biological chemistry High 7961908
1997 Alpha-SNAP C-terminal residues (including conserved Leu294) are required to stimulate NSF ATPase activity; stimulation of NSF ATPase by alpha-SNAP is required for 20S complex disassembly (VAMP dissociation) and for Ca2+-dependent exocytosis in chromaffin cells. Truncation/point mutagenesis of alpha-SNAP, in vitro ATPase assay, permeabilized chromaffin cell exocytosis assay The Journal of cell biology High 9362506
1995 NSF and alpha-SNAP are required for basolateral (but not apical) transport from TGN to plasma membrane in MDCK cells; a Rab-NSF-SNAP-SNARE mechanism operates in basolateral transport while the apical pathway is independent of this machinery. Streptolysin O-permeabilized MDCK cell transport assay with anti-NSF antibodies and recombinant proteins Cell High 7758111
1995 NSF together with SNAPs and the vesicle docking protein p115, as well as the NSF-like ATPase p97, are each sufficient to restore Golgi cisternal regrowth from mitotic Golgi fragments, but produce morphologically distinct cisternae, indicating distinct roles in Golgi reassembly. Cell-free Golgi reassembly assay with NEM inhibition, salt washing, and recombinant protein rescue Cell High 7553851
1998 Alpha-SNAP and syntaxin 5 are common components of both the NSF pathway and the p97 pathway for Golgi cisternal reassembly; p47 (a p97 cofactor) binds directly to syntaxin 5 and competes with alpha-SNAP for binding, playing an analogous adaptor role to alpha-SNAP for NSF. Cell-free Golgi reassembly assay, antibody inhibition, recombinant protein binding assays Cell High 9506515
1997 Yeast vacuole fusion requires Sec18p (NSF)/Sec17p (alpha-SNAP) for a symmetric 'priming' step preceding docking, and then requires the Rab GTPase Ypt7p for the docking step itself; priming produces a labile state that must be rapidly captured by docking. In vitro yeast vacuole fusion assay with affinity-purified antibodies and recombinant proteins, biochemical and microscopic docking assays The Journal of cell biology High 9015302
1996 Sec17p (alpha-SNAP) and Sec18p (NSF) are required for homotypic yeast vacuole fusion in vitro; vacuole-to-vacuole fusion is stimulated by fatty acyl-CoA compounds in a Sec18p-dependent fashion, and a cytosolic factor activates vacuole membrane-bound Sec18p. Cell-free yeast vacuole fusion assay with affinity-purified antibodies and recombinant proteins The EMBO journal High 8670830
1998 LMA1 (a heterodimer of thioredoxin and IB2) requires Sec18p for high-affinity binding to vacuoles; upon Sec18p ATP hydrolysis, LMA1 transfers to and stabilizes a Vam3p (t-SNARE) complex; LMA1 is subsequently released in a phosphatase-regulated reaction, coupling priming to t-SNARE stabilization. In vitro yeast vacuole fusion assay, affinity binding experiments, co-immunoprecipitation Cell High 9657146
1997 LMA1 heterodimer (thioredoxin + IB2) cooperates with Sec18p/NSF at an early priming step in vacuole fusion; Sec18p acts first and LMA1 stabilizes vacuoles after Sec17p/Sec18p action; LMA1 cannot act before Sec18p. In vitro yeast vacuole fusion with purified proteins, order-of-addition experiments The Journal of cell biology High 9015301
2001 Ergosterol is required for the Sec18p/NSF-mediated priming step of homotypic vacuole fusion; ergosterol ligands (filipin, nystatin, amphotericin B) block Sec17p release from vacuoles, and cholesterol/ergosterol supplementation stimulates in vitro fusion. In vitro yeast vacuole fusion assay, sterol addition/depletion, inhibitor studies The EMBO journal High 11483507
1998 NSF interacts directly and selectively with residues Lys-844 to Gln-853 of the GluR2 AMPA receptor C-terminal domain (with Asn-851 critical); this interaction requires all three NSF domains; loading a blocking decapeptide or anti-NSF antibody into hippocampal CA1 neurons causes a progressive decrement in AMPA receptor-mediated synaptic transmission. Direct binding assay, site-directed mutagenesis, intracellular peptide/antibody loading with electrophysiology Neuron High 9697854
1998 GluR2 C-terminal domain interacts with NSF and alpha- and beta-SNAPs; the GluR2-NSF-SNAP complex assembly is reversible and requires ATP hydrolysis, analogous to SNARE complex disassembly; NSF is proposed to act as a chaperone in AMPA receptor processing. GST pulldown, co-immunoprecipitation, ATP hydrolysis-dependent complex assay Neuron High 9697855
1999 Disruption of NSF-GluR2 interaction by intracellular infusion of blocking peptide (pep2m) into hippocampal neurons causes rapid decrease in AMPA receptor-mediated mEPSC frequency and reduces surface expression of GluR2-containing AMPA receptors, without affecting NMDA receptor surface expression. Intracellular peptide infusion, whole-cell electrophysiology (mEPSC recording), immunofluorescence surface labeling in cultured neurons Neuron High 10399941
2001 NSF-mediated SNARE complex disassembly occurs after synaptic vesicle fusion, not at the fusion step itself; in Drosophila comatose (NSF) mutants, blocking evoked fusion delays SNARE complex accumulation and paralysis, and the full vesicle pool can be depleted in NSF/shibire double mutants, showing NSF recycles SNARE proteins between exocytosis and endocytosis. Drosophila genetic analysis, temperature-sensitive paralysis, SNARE complex immunoblotting, double mutant epistasis Proceedings of the National Academy of Sciences of the United States of America High 11593041
1998 Injection of SNAP-stimulating peptides that inhibit NSF ATPase activity into squid giant presynaptic terminals reduces neurotransmitter release amount and slows its kinetics; this effect requires vesicle turnover and acts after vesicle docking, establishing NSF as a participant in regulating synaptic vesicle exocytosis kinetics. Intracellular peptide injection, squid giant synapse electrophysiology Science High 9469810
1998 In Drosophila comatose (dNSF-1) mutants at restrictive temperature, repetitive stimulation causes progressive activity-dependent reduction in neurotransmitter release and accumulation of docked vesicles at the active zone; NSF does not directly participate in vesicle-membrane fusion but maintains the pool of docked vesicles competent for calcium-triggered fusion (priming). Electrophysiology at Drosophila NMJ, transmission electron microscopy of synaptic terminals The Journal of neuroscience High 9852561
2000 Trans-SNARE complexes (SNAREpins) are functionally resistant to disruption by NSF and alpha-SNAP; this resistance is acquired at the moment SNAREpins form and commit to fusion, allowing fusion to proceed locally in a cellular environment that otherwise strongly favors SNARE disruption. In vitro reconstituted liposome fusion assay with NSF/alpha-SNAP addition The Journal of cell biology High 10831610
2002 NSF ATPase activity (with alpha-/beta-SNAPs) disassembles PICK1-GluR2 interactions: GluR2, PICK1, NSF, and SNAPs form a complex in the presence of ATPgammaS, and ATP hydrolysis by NSF disrupts PICK1-GluR2 interaction; this demonstrates a non-SNARE substrate for NSF disassembly activity and explains NSF-mediated synaptic stabilization of AMPARs. Co-immunoprecipitation, in vitro disassembly assay with ATPgammaS vs. ATP, neuronal overexpression with receptor trafficking readout Neuron High 11931741
2001 The ionic layer glutamine of syntaxin is required for efficient NSF/alpha-SNAP-mediated dissociation of the SNARE complex; when mutated, the complex still binds alpha-SNAP and NSF and is released upon ATP hydrolysis, but does not dissociate into monomers, indicating the syntaxin glutamine couples ATP hydrolysis to complex dissociation. Site-directed mutagenesis of syntaxin ionic layer, in vitro SNARE disassembly assay Proceedings of the National Academy of Sciences of the United States of America High 11762430
1998 NSF is a hexamer (not tetramer or trimer) stabilized by D2 domain oligomerization; demonstrated by sedimentation equilibrium and velocity analytical ultracentrifugation, transmission EM with rotational image analysis, scanning transmission EM, and multiangle light scattering. Analytical ultracentrifugation, EM, MALS — multiple biophysical techniques The Journal of biological chemistry High 9624162
2003 ATP-hydrolysis-deficient NSF(E329Q) (dominant negative) promotes Golgi stack disassembly into dispersed vesicles and inhibits intra-Golgi transport; in contrast, p97(E578Q) does not affect Golgi structure or transport, establishing that NSF but not p97 directly regulates membrane fusion at the Golgi. Dominant-negative ATPase mutant expression in mammalian cells, Golgi morphology, glycosaminoglycan sulfation assay, VSV-G transport Molecular biology of the cell High 14617820
2002 NSF disassembles Golgi SNAREs during mitotic Golgi fragmentation; a subsequent ATPase-independent NSF activity during reassembly catalyzes (with alpha-SNAP) the binding of GATE-16 to the v-SNARE GOS-28 in an ATP-but-not-hydrolysis-requiring manner, protecting the v-SNARE and regulating SNARE function for Golgi membrane fusion. Cell-free Golgi reassembly assay, NSF point mutants, immunoprecipitation, comatose mutant analysis The Journal of cell biology High 12070132
2015 NSF disassembles a single SNARE complex in one round of ATP turnover via a 'spring-loaded' mechanism: ATP cleavage builds internal tension within the NSF hexamer following phosphate dissociation; after a latent period of tens of seconds, NSF releases built-up tension in a burst within 20 ms, resulting in SNARE disassembly and release. Single-molecule fluorescence spectroscopy and magnetic tweezers Science High 25814585
2018 Cryo-EM structure of NSF/2×alpha-SNAP/neuronal SNARE complex (20S supercomplex) at ~3.9 Å reveals two alpha-SNAP molecules interfacing with a specific surface of the SNARE complex via electrostatic interactions; 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 NSF tyrosine and SNAP-25A backbone atoms, likely preceding ATP hydrolysis. Electron cryo-microscopy structure determination at near-atomic resolution eLife High 30198481
2016 LRRK2 phosphorylates NSF at Thr-645 in the ATP binding pocket of the D2 domain; phosphorylated NSF displays enhanced ATPase activity and increased rate of SNARE complex disassembly; Thr645Ala substitution abrogates LRRK2-mediated enhanced ATPase activity. In vitro kinase assay, mass spectrometry phosphosite identification, site-directed mutagenesis, ATPase activity assay, SNARE disassembly assay Molecular neurodegeneration High 26758690
1999 Beta-arrestin1 interacts with NSF in a conformation-dependent manner, preferentially binding the ATP-bound form; NSF overexpression enhances agonist-mediated beta2-adrenergic receptor internalization and rescues inhibition caused by a dominant-negative beta-arrestin1 phosphomimetic. Yeast two-hybrid, in vitro pulldown with recombinant proteins, co-immunoprecipitation from cells, overexpression in HEK293 cells with receptor internalization assay The Journal of biological chemistry High 10196135
2012 Single-particle cryo-EM reconstruction of Chinese hamster NSF hexamer in ATPgammaS, ADP-AlFx, and ADP states, and of the 20S particle, reveals parallel arrangement of D1 and D2 domains, nucleotide-dependent conformational changes, and two interaction interfaces between NSF/SNAP and the SNARE complex (C terminus and N-terminal half of the SNARE bundle). Single-particle cryo-EM and negative stain EM, 3D reconstruction Nature structural & molecular biology High 22307055
1999 NSF-GluR2 interaction blockade (pep2m peptide) in hippocampal CA1 neurons prevents homosynaptic LTD induction; LTD saturation prevents pep2m-induced reduction in AMPAR EPSCs, indicating that the NSF-dependent pool of AMPARs is the same pool removed during LTD expression. Intracellular peptide infusion, hippocampal slice electrophysiology, minimal stimulation experiments Neuron High 10571232
2002 NSF binding to GluR2 is required to prevent excess AMPA receptor internalization in response to AMPA or NMDA; GluR2 mutants lacking NSF binding undergo greater stimulus-induced endocytosis. GRIP/ABP binding stabilizes an intracellular pool of internalized AMPARs, inhibiting their recycling. Epitope-tagged GluR2 mutants in hippocampal neurons, surface biotinylation, AMPA/NMDA-induced internalization assay Proceedings of the National Academy of Sciences of the United States of America High 12011465
2002 AP2 clathrin adaptor associates with a GluR2 region overlapping the NSF binding site; AP2 is specifically required for NMDA receptor-induced (not ligand-dependent) AMPA receptor internalization and hippocampal LTD; NSF maintains basal synaptic AMPA receptor responses but is not directly required for NMDA receptor-mediated internalization or LTD. GluR2 mutants dissociating NSF vs. AP2 binding, co-immunoprecipitation, peptide competition, hippocampal slice LTD Neuron High 12441055
2010 Polo-like kinase 2 (Plk2) directly interacts with NSF through a motif independent of canonical polo box sites; Plk2 disrupts NSF-GluR2 interaction independently of its kinase activity, promoting loss of surface GluR2, greater GluR2 association with PICK1/GRIP1, and decreased synaptic AMPAR current during chronic overexcitation. Co-immunoprecipitation, surface biotinylation, whole-cell patch clamp in rat hippocampal neurons, dominant-negative and kinase-dead mutants Nature neuroscience High 20802490
2010 NSF binding to the GluR2 C-terminal domain (via the NSF binding site) is required for plasma membrane insertion of GluR2; RNA editing of the Q/R site also regulates plasma membrane insertion; visualized by pHluorin-TIRF microscopy. pHluorin-tagged GluR2 TIRF microscopy, NSF-binding site mutant, Q/R editing mutant Proceedings of the National Academy of Sciences of the United States of America High 20534470
2001 GABARAP (a GABA-A receptor gamma2 subunit-binding protein) directly binds NSF; GABARAP-NSF complexes are detected in neurons and the two proteins colocalize in intracellular membrane compartments (Golgi and postsynaptic cisternae), suggesting GABARAP links GABA-A receptor transport to NSF-mediated trafficking. GST pulldown, co-immunoprecipitation from brain, immunofluorescence colocalization Molecular and cellular neurosciences Medium 11461150
2006 NSF directly interacts with the GABA-B receptor (GBR) heterodimer in rat brain synaptosomes and CHO cells; a peptide (TAT-Pep-27) blocking NSF-GBR2 interaction abolishes agonist-promoted GBR desensitization and prevents PKC recruitment and receptor phosphorylation, without affecting basal signaling or receptor internalization. Co-immunoprecipitation from synaptosomes and CHO cells, peptide inhibitor, hippocampal slice electrophysiology, PKC recruitment assay The EMBO journal High 16724110
2005 GluR2-NSF interaction is required for rapid direct insertion of AMPA receptors into synaptic (not extrasynaptic) plasma membrane; introducing the NSF binding site into GluR3 confers GluR2-like kinetics and synaptic insertion, demonstrating sufficiency of NSF interaction for synaptic targeting. Cell-surface thrombin cleavage assay, live-cell immunostaining in hippocampal neurons, GluR2/GluR3 chimeric constructs Molecular and cellular neurosciences High 15797712
2005 NSF and PICK1 are both required for calcium-permeable AMPA receptor plasticity (CARP); PICK1 (but not NSF) regulates formation of extrasynaptic GluR2-containing receptor pools that are laterally mobilized into synapses during CARP. Peptide inhibitors, dominant-negative constructs, electrophysiology in hippocampal neurons Neuron High 15797551
2009 Alpha-SNAP contains a conserved membrane attachment site in an N-terminal extended loop (including two conserved phenylalanine residues); mutation of these residues prevents alpha-SNAP from facilitating disassembly of membrane-bound (but not soluble) SNARE complexes, indicating the disassembly machinery is adapted to attack membrane-bound SNARE complexes. In vitro SNARE disassembly assay comparing soluble and membrane-bound substrates, site-directed mutagenesis The Journal of biological chemistry High 19762473
2018 NSF disassembles parallel ternary SNARE complexes in a single step within 100 ms; short-lived disassembled states represent failed disassembly or immediate reassembly; complexin-1 competes with alpha-SNAP for SNARE complex binding, reducing disassembly rate similar to decreased alpha-SNAP concentration, potentially differentially regulating cis vs. trans SNARE complex disassembly. Single-molecule FRET assay monitoring repeated rounds of NSF disassembly/reassembly eLife High 29985126
2019 Munc18-1 and Munc13-1 are required for trans-SNARE complex formation in the presence of NSF-alpha-SNAP; Munc18-1, Munc13-1, complexin-1, and synaptotagmin-1 all contribute to maintaining assembled trans-SNARE complexes against NSF-alpha-SNAP-mediated disassembly (preventing de-priming). In vitro reconstituted trans-SNARE complex formation assay with purified proteins and NSF-alpha-SNAP eLife High 30657450
1999 Alpha-SNAP/NSF function is required at an early priming step in chromaffin cell exocytosis that increases the amplitude of both the exocytotic burst and the slow (recruitment) component but does not alter fusion kinetics of the readily releasable pool; NEM partially inhibits the slow component without affecting the burst. Capacitance measurement and electrochemical amperometry in chromaffin cells, flash photolysis of caged Ca2+, recombinant alpha-SNAP and mutants The EMBO journal High 10369670
2006 Loss of nsf in zebrafish causes defects in myelin basic protein expression and in localization of sodium channel proteins at nodes of Ranvier; chimeric analysis shows nsf acts cell-autonomously in neurons for sodium channel clustering; this role is independent of nsf function in synaptic vesicle fusion. Zebrafish forward genetic screen, chimeric larvae, pharmacological analysis, immunofluorescence Current biology High 16581508
2002 In Dictyostelium, temperature-sensitive NSF (nsfA) mutants lose cell polarity, chemotaxis, and membrane recycling (macropinocytosis, phagocytosis, FM1-43 internalization) at restrictive temperature while retaining cAMP-induced actin responses, demonstrating NSF-catalysed membrane recycling is required for maintenance of cell polarity and locomotion. Temperature-sensitive mutagenesis, FITC-dextran uptake, FM1-43 internalization, live-cell imaging of chemotaxis Development High 12183371
1999 Alpha-SNAP and NSF are required for a prefusion priming step in human sperm acrosome reaction (AR), acting after acrosome tethering to plasma membrane but before intra-acrosomal Ca2+ efflux; alpha-SNAP exerts its effect through interaction with NSF. Streptolysin O-permeabilized human sperm, anti-alpha-SNAP antibody inhibition, recombinant alpha-SNAP addition, Western blot and immunostaining Molecular human reproduction High 15542541
1999 NSF is required for platelet granule (alpha-granule and dense-granule) secretion; NSF-sequence-mimicking peptides that inhibit alpha-SNAP-stimulated NSF ATPase activity inhibit secretion in permeabilized human platelets; anti-NSF antibodies also inhibit secretion, rescued by recombinant NSF. Permeabilized human platelet secretion assay, peptide and antibody inhibition, recombinant NSF rescue Blood High 10438719
1998 NSF and alpha-/beta-SNAPs mediate dissociation of the GS28-syntaxin 5 Golgi SNARE complex; this dissociation requires ATP hydrolysis by NSF and a concerted action of both alpha-SNAP and NSF; GS28 (not syntaxin 5) binds directly to immobilized alpha-SNAP after complex dissociation. Co-immunoprecipitation from Golgi extracts, in vitro disassembly assay with recombinant proteins, ATP/ATPgammaS conditions The Journal of biological chemistry High 9325254
2005 NSF/SNAPs mediate the first fusion reaction in ER network reformation from mitotic fragments (generating connecting intermediates), while p97/p47/VCIP135 mediates a subsequent fusion; both processes involve the t-SNARE syntaxin 18. SLO-permeabilized CHO cell ER reformation assay, antibody and recombinant protein inhibition, GFP-HSP47 live imaging Genes to cells High 16164599

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1997 Structure and conformational changes in NSF and its membrane receptor complexes visualized by quick-freeze/deep-etch electron microscopy. Cell 664 9267032
1990 SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast. Cell 459 2111733
1998 NSF binding to GluR2 regulates synaptic transmission. Neuron 452 9697854
1991 Compartmental organization of Golgi-specific protein modification and vacuolar protein sorting events defined in a yeast sec18 (NSF) mutant. The Journal of cell biology 376 2071670
2002 Clathrin adaptor AP2 and NSF interact with overlapping sites of GluR2 and play distinct roles in AMPA receptor trafficking and hippocampal LTD. Neuron 344 12441055
1995 An NSF-like ATPase, p97, and NSF mediate cisternal regrowth from mitotic Golgi fragments. Cell 316 7553851
1998 The AMPA receptor GluR2 C terminus can mediate a reversible, ATP-dependent interaction with NSF and alpha- and beta-SNAPs. Neuron 306 9697855
1995 Endothelial caveolae have the molecular transport machinery for vesicle budding, docking, and fusion including VAMP, NSF, SNAP, annexins, and GTPases. The Journal of biological chemistry 300 7782301
2001 Autophagosome requires specific early Sec proteins for its formation and NSF/SNARE for vacuolar fusion. Molecular biology of the cell 297 11694599
1990 An abundant and ubiquitous homo-oligomeric ring-shaped ATPase particle related to the putative vesicle fusion proteins Sec18p and NSF. The EMBO journal 261 2140770
1999 Surface expression of AMPA receptors in hippocampal neurons is regulated by an NSF-dependent mechanism. Neuron 254 10399941
1997 SNAREs and NSF in targeted membrane fusion. Current opinion in cell biology 249 9261050
1999 Hippocampal LTD expression involves a pool of AMPARs regulated by the NSF-GluR2 interaction. Neuron 240 10571232
1993 SNAP family of NSF attachment proteins includes a brain-specific isoform. Nature 240 8455721
1998 Syntaxin 5 is a common component of the NSF- and p97-mediated reassembly pathways of Golgi cisternae from mitotic Golgi fragments in vitro. Cell 224 9506515
2005 Calcium-permeable AMPA receptor plasticity is mediated by subunit-specific interactions with PICK1 and NSF. Neuron 212 15797551
1997 Docking of yeast vacuoles is catalyzed by the Ras-like GTPase Ypt7p after symmetric priming by Sec18p (NSF). The Journal of cell biology 207 9015302
1995 Different requirements for NSF, SNAP, and Rab proteins in apical and basolateral transport in MDCK cells. Cell 205 7758111
2001 The subcellular distribution of GABARAP and its ability to interact with NSF suggest a role for this protein in the intracellular transport of GABA(A) receptors. Molecular and cellular neurosciences 190 11461150
1988 Characterization of a component of the yeast secretion machinery: identification of the SEC18 gene product. Molecular and cellular biology 188 3054509
1998 Fusion of lysosomes with late endosomes produces a hybrid organelle of intermediate density and is NSF dependent. The Journal of cell biology 170 9456319
2002 NSF ATPase and alpha-/beta-SNAPs disassemble the AMPA receptor-PICK1 complex. Neuron 165 11931741
1993 Inactivation of YME1, a member of the ftsH-SEC18-PAS1-CDC48 family of putative ATPase-encoding genes, causes increased escape of DNA from mitochondria in Saccharomyces cerevisiae. Molecular and cellular biology 164 8355690
1996 Homotypic vacuole fusion requires Sec17p (yeast alpha-SNAP) and Sec18p (yeast NSF). The EMBO journal 156 8670830
2003 Distinct roles for the AAA ATPases NSF and p97 in the secretory pathway. Molecular biology of the cell 152 14617820
1997 Stimulation of NSF ATPase activity by alpha-SNAP is required for SNARE complex disassembly and exocytosis. The Journal of cell biology 147 9362506
1997 A novel Sec18p/NSF-dependent complex required for Golgi-to-endosome transport in yeast. Molecular biology of the cell 142 9201718
2004 A library of 7TM receptor C-terminal tails. Interactions with the proposed post-endocytic sorting proteins ERM-binding phosphoprotein 50 (EBP50), N-ethylmaleimide-sensitive factor (NSF), sorting nexin 1 (SNX1), and G protein-coupled receptor-associated sorting protein (GASP). The Journal of biological chemistry 139 15452121
2001 Ergosterol is required for the Sec18/ATP-dependent priming step of homotypic vacuole fusion. The EMBO journal 135 11483507
1995 Clostridial neurotoxins compromise the stability of a low energy SNARE complex mediating NSF activation of synaptic vesicle fusion. The EMBO journal 125 7588600
1999 Identification of NSF as a beta-arrestin1-binding protein. Implications for beta2-adrenergic receptor regulation. The Journal of biological chemistry 117 10196135
2002 Differential roles for NSF and GRIP/ABP in AMPA receptor cycling. Proceedings of the National Academy of Sciences of the United States of America 108 12011465
2016 LRRK2 phosphorylates pre-synaptic N-ethylmaleimide sensitive fusion (NSF) protein enhancing its ATPase activity and SNARE complex disassembling rate. Molecular neurodegeneration 107 26758690
2007 Cellular functions of NSF: not just SNAPs and SNAREs. FEBS letters 103 17397838
2001 SNARE-complex disassembly by NSF follows synaptic-vesicle fusion. Proceedings of the National Academy of Sciences of the United States of America 100 11593041
2000 SNAREpins are functionally resistant to disruption by NSF and alphaSNAP. The Journal of cell biology 99 10831610
1994 The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. The Journal of biological chemistry 98 7961908
2001 N-ethylmaleimide sensitive factor (NSF) structure and function. International review of cytology 97 11352269
1995 A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells. The EMBO journal 97 7835334
1998 Analysis of regulated exocytosis in adrenal chromaffin cells: insights into NSF/SNAP/SNARE function. BioEssays : news and reviews in molecular, cellular and developmental biology 93 9619104
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1998 Synaptic physiology and ultrastructure in comatose mutants define an in vivo role for NSF in neurotransmitter release. The Journal of neuroscience : the official journal of the Society for Neuroscience 88 9852561
1999 Early requirement for alpha-SNAP and NSF in the secretory cascade in chromaffin cells. The EMBO journal 85 10369670
1998 Regulation of neurotransmitter release kinetics by NSF. Science (New York, N.Y.) 85 9469810
2001 Association of a novel PDZ domain-containing peripheral Golgi protein with the Q-SNARE (Q-soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor) protein syntaxin 6. The Journal of biological chemistry 84 11384996
2015 Spring-loaded unraveling of a single SNARE complex by NSF in one round of ATP turnover. Science (New York, N.Y.) 81 25814585
1998 A revised model for the oligomeric state of the N-ethylmaleimide-sensitive fusion protein, NSF. The Journal of biological chemistry 77 9624162
2018 Structural principles of SNARE complex recognition by the AAA+ protein NSF. eLife 75 30198481
1998 LMA1 binds to vacuoles at Sec18p (NSF), transfers upon ATP hydrolysis to a t-SNARE (Vam3p) complex, and is released during fusion. Cell 74 9657146
2016 Disease resistance through impairment of α-SNAP-NSF interaction and vesicular trafficking by soybean Rhg1. Proceedings of the National Academy of Sciences of the United States of America 73 27821740
2010 Plasma membrane insertion of the AMPA receptor GluA2 subunit is regulated by NSF binding and Q/R editing of the ion pore. Proceedings of the National Academy of Sciences of the United States of America 72 20534470
2008 N-ethylmaleimide-sensitive fusion protein (NSF) is involved in central sensitization in the spinal cord through GluR2 subunit composition switch after inflammation. The European journal of neuroscience 71 18598260
2002 Sequential SNARE disassembly and GATE-16-GOS-28 complex assembly mediated by distinct NSF activities drives Golgi membrane fusion. The Journal of cell biology 70 12070132
2019 Multiple factors maintain assembled trans-SNARE complexes in the presence of NSF and αSNAP. eLife 67 30657450
2003 NSF and p97/VCP: similar at first, different at last. FEBS letters 67 14630332
1996 Characterization and subcellular localization of target membrane soluble NSF attachment protein receptors (t-SNAREs) in macrophages. Syntaxins 2, 3, and 4 are present on phagosomal membranes. Journal of immunology (Baltimore, Md. : 1950) 66 8666810
1996 Reconstitution of transcytosis in SLO-permeabilized MDCK cells: existence of an NSF-dependent fusion mechanism with the apical surface of MDCK cells. The EMBO journal 65 8612570
1994 Neurally expressed Drosophila genes encoding homologs of the NSF and SNAP secretory proteins. Proceedings of the National Academy of Sciences of the United States of America 62 8202553
1999 Cytosolic ATPases, p97 and NSF, are sufficient to mediate rapid membrane fusion. The EMBO journal 61 10205162
1997 A heterodimer of thioredoxin and I(B)2 cooperates with Sec18p (NSF) to promote yeast vacuole inheritance. The Journal of cell biology 61 9015301
2010 Plk2 attachment to NSF induces homeostatic removal of GluA2 during chronic overexcitation. Nature neuroscience 60 20802490
2008 Circulating fibrocytes: cellular basis for NSF. Journal of the American College of Radiology : JACR 60 18180007
2017 Characterization of the Soluble NSF Attachment Protein gene family identifies two members involved in additive resistance to a plant pathogen. Scientific reports 59 28338077
2009 A conserved membrane attachment site in alpha-SNAP facilitates N-ethylmaleimide-sensitive factor (NSF)-driven SNARE complex disassembly. The Journal of biological chemistry 56 19762473
2004 alpha-SNAP and NSF are required in a priming step during the human sperm acrosome reaction. Molecular human reproduction 56 15542541
1996 Domains of alpha-SNAP required for the stimulation of exocytosis and for N-ethylmalemide-sensitive fusion protein (NSF) binding and activation. Molecular biology of the cell 55 8744944
2005 SEC18/NSF-independent, protein-sorting pathway from the yeast cortical ER to the plasma membrane. The Journal of cell biology 53 15911878
2016 Review: Progresses in understanding N-ethylmaleimide sensitive factor (NSF) mediated disassembly of SNARE complexes. Biopolymers 51 27062050
2015 Sec17 can trigger fusion of trans-SNARE paired membranes without Sec18. Proceedings of the National Academy of Sciences of the United States of America 51 25902545
2011 Requirements for the catalytic cycle of the N-ethylmaleimide-Sensitive Factor (NSF). Biochimica et biophysica acta 51 21689688
1998 Soluble NSF-attachment proteins. The international journal of biochemistry & cell biology 51 9693958
1999 Constitutive calcium-independent release of Toxoplasma gondii dense granules occurs through the NSF/SNAP/SNARE/Rab machinery. The Journal of biological chemistry 50 9891012
1995 Distinct roles for N-ethylmaleimide-sensitive fusion protein (NSF) suggested by the identification of a second Drosophila NSF homolog. The Journal of biological chemistry 48 7642522
2015 Recent Advances in Deciphering the Structure and Molecular Mechanism of the AAA+ ATPase N-Ethylmaleimide-Sensitive Factor (NSF). Journal of molecular biology 47 26546278
2005 NSF/SNAPs and p97/p47/VCIP135 are sequentially required for cell cycle-dependent reformation of the ER network. Genes to cells : devoted to molecular & cellular mechanisms 46 16164599
2001 The ionic layer is required for efficient dissociation of the SNARE complex by alpha-SNAP and NSF. Proceedings of the National Academy of Sciences of the United States of America 46 11762430
1992 The activity of Golgi transport vesicles depends on the presence of the N-ethylmaleimide-sensitive factor (NSF) and a soluble NSF attachment protein (alpha SNAP) during vesicle formation. The Journal of cell biology 46 1522110
2017 Sec17/Sec18 act twice, enhancing membrane fusion and then disassembling cis-SNARE complexes. eLife 45 28718762
2012 Structural characterization of full-length NSF and 20S particles. Nature structural & molecular biology 45 22307055
2006 Coordinated action of NSF and PKC regulates GABAB receptor signaling efficacy. The EMBO journal 43 16724110
1999 A critical role for N-ethylmaleimide-sensitive fusion protein (NSF) in platelet granule secretion. Blood 43 10438719
2008 Gadolinium-containing MRI contrast agents: important variations on a theme for NSF. Journal of the American College of Radiology : JACR 42 18180006
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1998 NSF--fusion and beyond. Trends in cell biology 41 9861668
1996 Association of the fusion protein NSF with clathrin-coated vesicle membranes. The EMBO journal 41 8631296
2006 nsf is essential for organization of myelinated axons in zebrafish. Current biology : CB 40 16581508
2018 NSF-mediated disassembly of on- and off-pathway SNARE complexes and inhibition by complexin. eLife 39 29985126
1994 Identification of a novel mammalian member of the NSF/CDC48p/Pas1p/TBP-1 family through heterologous expression in yeast. FEBS letters 39 8082782
2008 NSF, Unc-18-1, dynamin-1 and HSP90 are inclusion body components in neuronal intranuclear inclusion disease identified by anti-SUMO-1-immunocapture. Acta neuropathologica 38 18836734
2002 Cell polarity and locomotion, as well as endocytosis, depend on NSF. Development (Cambridge, England) 37 12183371
1995 Reconstitution of vesiculated Golgi membranes into stacks of cisternae: requirement of NSF in stack formation. The Journal of cell biology 37 7730397
1975 T cell-dependent mediator in the immune response. III. The role of non-specific factor (NSF) in the in vitro immune response. Immunology 37 1092612
2005 NSF interaction is important for direct insertion of GluR2 at synaptic sites. Molecular and cellular neurosciences 36 15797712
2011 Both pre- and postsynaptic activity of Nsf prevents degeneration of hair-cell synapses. PloS one 35 22073277
1997 N-Ethylmaleimide-sensitive factor (NSF) and alpha-soluble NSF attachment proteins (SNAP) mediate dissociation of GS28-syntaxin 5 Golgi SNAP receptors (SNARE) complex. The Journal of biological chemistry 34 9325254
2002 NSF regulates membrane traffic along multiple pathways in Paramecium. Journal of cell science 33 12244131
2017 Roles of Cellular NSF Protein in Entry and Nuclear Egress of Budded Virions of Autographa californica Multiple Nucleopolyhedrovirus. Journal of virology 32 28747507
2001 Partitioning of N-ethylmaleimide-sensitive fusion (NSF) protein function in Drosophila melanogaster: dNSF1 is required in the nervous system, and dNSF2 is required in mesoderm. Genetics 32 11333235
1991 Structure of Saccharomyces cerevisiae alg3, sec18 mutant oligosaccharides. The Journal of biological chemistry 32 2005096
2016 Dysferlin Binds SNAREs (Soluble N-Ethylmaleimide-sensitive Factor (NSF) Attachment Protein Receptors) and Stimulates Membrane Fusion in a Calcium-sensitive Manner. The Journal of biological chemistry 31 27226605