| 1995 |
VAMP-2 is essential for Ca2+-dependent insulin secretion in pancreatic beta-cells. Tetanus toxin and botulinum B neurotoxin selectively cleaved VAMP-2 and cellubrevin in permeabilized beta-cells and abolished Ca2+-induced insulin release (IC50 ~15 nM), but did not prevent GTP-gamma-S-stimulated secretion, demonstrating that VAMP-2 selectively controls the Ca2+-mediated exocytotic pathway. |
Tetanus toxin and botulinum B neurotoxin cleavage assay in streptolysin-O permeabilized insulin-secreting cells; subcellular fractionation; confocal microscopy |
The EMBO journal |
High |
7796801
|
| 1997 |
VAMP2 functions as a vesicle-SNARE (v-SNARE) required for insulin-stimulated GLUT4 translocation to the plasma membrane in adipocytes. Expression of the cytoplasmic domain of VAMP2 inhibited insulin-stimulated GLUT4 but not GLUT1 translocation, and immunoprecipitation of syntaxin 4 cytoplasmic domain co-precipitated GLUT4-containing vesicles in an insulin-stimulated manner. |
Recombinant vaccinia virus expression; single-cell microinjection; co-immunoprecipitation |
Molecular and cellular biology |
High |
9111311
|
| 1996 |
VAMP-2 co-localizes with GLUT-4 in a specialized post-endosomal intracellular compartment in adipocytes that is segregated from recycling endosomes (transferrin receptor/cellubrevin-positive). Endosomal ablation with transferrin-HRP/DAB removed >90% of cellubrevin but spared 90% of VAMP-2, and immunoisolated GLUT-4 vesicles contained VAMP-2. |
Endosomal ablation (Tf-HRP/DAB), immunoadsorption of vesicles, immuno-electron microscopy, subcellular fractionation |
The Journal of cell biology |
High |
8707843
|
| 1995 |
VAMP2 is expressed on AQP2-containing intracellular vesicles in kidney collecting duct principal cells, as demonstrated by co-immunoprecipitation and double immunolabeling; VAMP2 in kidney membranes is cleaved by tetanus toxin, establishing it as a tetanus-toxin-sensitive SNARE in this tissue. Quantitative immuno-EM showed highly significant co-localization of AQP2 and VAMP2 on the same vesicles (P<0.0001). |
Immunoblotting, immunoisolation with anti-VAMP2 antibodies, double immunolabeling immuno-EM, tetanus toxin cleavage |
The Journal of clinical investigation |
High |
7560075
|
| 1998 |
VAMP-2 forms binary, saturable interactions with both syntaxin-4 and SNAP23 in vitro, establishing the non-neuronal SNARE complex for exocytosis. Unlike neuronal SNAP25, SNAP23 did not potentiate VAMP-2 binding to syntaxin-4. PKA phosphorylation of syntaxin-4 (but not casein kinase II) disrupted its binding to SNAP23, indicating phosphorylation-dependent regulation of SNARE complex formation. |
In vitro binary binding assay; phosphorylation by exogenous kinases (PKA, CKII, PKC); phosphate incorporation quantification |
Biochemistry |
High |
9693005
|
| 1997 |
A dual Rab/VAMP2 receptor protein (PRA1) was identified that specifically binds VAMP2 (but not VAMP1 or cellubrevin); this specificity requires the proline-rich domain and transmembrane domain of VAMP2. The interaction is distinct from VAMP2 binding to syntaxin or syntaxin/SNAP-25, suggesting PRA1 links Rab GTPases and VAMP2 in vesicle docking/fusion control. |
Yeast two-hybrid screen; in vitro binding assays; chimeric/deletion mutant analysis |
The Journal of biological chemistry |
High |
9341137
|
| 2000 |
VAMP2, but not VAMP3/cellubrevin, specifically mediates insulin-dependent GLUT4 translocation in L6 myoblasts. Toxin-resistant VAMP2 rescued tetanus toxin-inhibited GLUT4 translocation, whereas toxin-resistant VAMP3 did not. Insulin caused cortical actin reorganization in which GLUT4 and VAMP2 (but not VAMP3) were clustered. |
Tetanus toxin light chain transfection; rescue with toxin-resistant VAMP2/3 constructs; single-cell fluorescence GLUT4 translocation assay; immunofluorescence |
Molecular biology of the cell |
High |
10888677
|
| 2000 |
SNAP23 mediates insulin-dependent GLUT4 translocation by acting as a bridging molecule between syntaxin4 (t-SNARE) and VAMP2 (v-SNARE) in 3T3-L1 adipocytes. A SNAP23 mutant (DeltaC8) that bound syntaxin4 but not VAMP2 and failed to mediate the syntaxin4-VAMP2 interaction acted as a dominant negative, blocking insulin-induced GLUT4 translocation. |
Adenovirus-mediated overexpression of SNAP23 mutants; in vitro and in vivo co-immunoprecipitation; GLUT4 translocation assay |
The Journal of biological chemistry |
High |
10713150
|
| 2003 |
Synaptophysin I (SypI) controls the targeting of VAMP2 to synaptic vesicles through direct protein-protein interaction requiring the cytoplasmic domain of VAMP2. Without SypI co-expression, VAMP2 travels in vesicles that constitutively fuse with the plasma membrane; co-expression of SypI restores correct sorting to SVs in a dose-dependent and VAMP2-specific manner. |
Fluorescent chimera expression in hippocampal neurons; live imaging; co-immunoprecipitation; dose-response analysis |
Molecular biology of the cell |
High |
14528015
|
| 2004 |
VAMP2-dependent exocytosis regulates plasma membrane insertion of TRPC3 channels and contributes to agonist-stimulated Ca2+ influx. TRPC3 N-terminus directly interacts with VAMP2; cleavage of VAMP2 by tetanus toxin reduced TRPC3 surface expression and decreased carbachol- and OAG-stimulated (but not thapsigargin-stimulated) Ca2+ influx. |
Co-immunoprecipitation; GFP-TRPC3 live imaging; FRAP; tetanus toxin cleavage; Ca2+ influx measurement |
Molecular cell |
High |
15327778
|
| 2006 |
The 1.65 Å crystal structure of BoNT/D light chain provides molecular details of how VAMP-2 is recognized and cleaved. A hydrophobic pocket recognizes the P1' VAMP-2 residue Leu60, and a second remote site recognizes the V1 SNARE motif. Comparison with BoNT/F-LC (which cleaves VAMP-2 one residue away) showed that BoNT/D uses hydrophobic interactions for V1 motif recognition whereas BoNT/F adopts a hydrophilic strategy. |
X-ray crystallography at 1.65 Å resolution; structural comparison |
Biochemistry |
High |
16519520
|
| 2007 |
CALM (clathrin assembly lymphoid myeloid leukemia protein) facilitates endocytosis of VAMP2 from the plasma membrane via its ANTH domain. CALM overexpression reduced surface VAMP2; CALM knockdown by RNAi accumulated surface VAMP2; the ANTH domain alone acted as a dominant-negative, establishing CALM as a specific endocytic adaptor for VAMP2. |
Overexpression; RNA interference knockdown; dominant-negative ANTH domain expression; cell surface VAMP2 quantification |
Traffic (Copenhagen, Denmark) |
High |
18182011
|
| 2007 |
Synaptophysin I specifies the exocytic pathway of VAMP2 by directing its sorting to vesicles before surface delivery, not by inhibiting VAMP2 endocytosis. Physical interaction between SypI and VAMP2 is required, mediated by the C-terminal domain of SypI. Dynamin and alpha-SNAP mutants blocking trafficking at the plasma membrane did not abolish SypI's effect on VAMP2 sorting. |
Ectopic expression in non-neuronal cells; co-immunoprecipitation; dominant-negative dynamin and alpha-SNAP mutants; fluorescence microscopy |
The Biochemical journal |
Medium |
17331077
|
| 2007 |
ProF (WD-repeat-propeller-FYVE protein) interacts with VAMP2 and PKCzeta, forming a ternary complex on vesicular membranes. VAMP2 can be phosphorylated by activated PKCzeta in vitro, and ProF acts as an adaptor that increases PKCzeta-dependent phosphorylation of VAMP2 in vitro. |
Co-immunoprecipitation of endogenous and overexpressed proteins; in vitro kinase assay; co-localization imaging |
The FEBS journal |
Medium |
17313651
|
| 2008 |
VAMP2 directly interacts with the T1 domain of the N-terminus of Kv2.1 potassium channel and enhances channel inactivation. This interaction was demonstrated in brain membranes and characterized by electrophysiology and in vitro binding; a chimeric Kv1.5N/Kv2.1 channel confirmed the N-terminal T1 domain requirement. |
In vitro binding assay; electrophysiology in Xenopus oocytes; co-immunoprecipitation from brain membranes; chimeric channel analysis; protein modeling |
Pflugers Archiv : European journal of physiology |
High |
18542995
|
| 2008 |
Munc18c directly inhibits bilayer fusion mediated by the syntaxin4/SNAP23/VAMP2 SNARE complex in a reconstituted liposome fusion assay, making contacts with both t-SNARE (syntaxin4) and v-SNARE (VAMP2) components. |
Reconstituted liposome fusion assay; biochemical binding characterization |
PloS one |
High |
19116655
|
| 2001 |
PKCzeta, activated by insulin, associates specifically with GLUT4 compartments and induces serine phosphorylation of the GLUT4-compartment-associated VAMP2. Dominant-negative PKCzeta disrupted GLUT4 compartment integrity and abrogated insulin-induced GLUT4 translocation, while active PKCzeta overexpression caused GLUT4 translocation in the absence of insulin. |
Adenovirus-mediated overexpression of wild-type and dominant-negative PKCzeta; subcellular fractionation; phosphoserine immunoprecipitation; glucose transport assay |
Molecular and cellular biology |
Medium |
11604519
|
| 2009 |
The t-SNAREs syntaxin4 and SNAP23 are required for tethering of GLUT4 vesicles to the plasma membrane, whereas v-SNARE VAMP2 is not required for tethering but is essential for the subsequent membrane fusion event. RNAi depletion of VAMP2 inhibited fusion without affecting tethering, while depletion of syntaxin4 or SNAP23 impaired tethering. |
RNAi knockdown in 3T3-L1 adipocytes; TIRF microscopy-based vesicle tethering and fusion assay |
Biochemical and biophysical research communications |
High |
20006577
|
| 2009 |
The accessory alpha-helix of complexin I (residues 27-48) can locally displace VAMP2 from the C-terminus of the SNARE four-helix bundle, making the complex weaker, but remains detached when the N-terminal region of complexin I (residues 1-26) is present. This suggests the balance between the accessory helix and N-terminal domain determines stimulatory vs. inhibitory complexin function. |
EPR spectroscopy of complexin-SNARE quaternary complex; site-directed spin labeling |
Journal of molecular biology |
Medium |
20026076
|
| 2011 |
VAMP2 (but not VAMP3) mediates cAMP-stimulated renin release from juxtaglomerular cells. VAMP2 co-localizes with renin-containing granules, and shRNA silencing of VAMP2 blocked cAMP-induced renin release by ~50% and impaired cAMP-stimulated exocytosis (FM1-43 assay), while VAMP3 silencing had no effect. |
Confocal colocalization; adenoviral shRNA knockdown; FM1-43 exocytosis assay; renin secretion measurement; tetanus toxin cleavage |
The Journal of biological chemistry |
High |
21708949
|
| 2013 |
Native alpha-synuclein promotes clustering of synaptic-vesicle mimics through specific interactions with both synaptobrevin-2/VAMP2 and anionic lipids. The lipid-binding-deficient A30P mutant disrupted clustering, while other familial PD mutants did not. Alpha-synuclein had little effect on Ca2+-triggered fusion in this reconstituted single-vesicle system. |
Single-vesicle optical microscopy; recombinant and native alpha-synuclein purified from mouse brain; vesicle clustering assay |
eLife |
High |
23638301
|
| 2013 |
Heat shock factor 1 (HSF1) transcriptional activity mediates alcohol-induced upregulation of Vamp2 (but not Vamp1) expression in mouse cortical neurons. This increased VAMP2 expression leads to increased frequency of GABAergic miniature IPSCs via HSF1, without affecting mEPSCs, establishing a specific presynaptic adaptation in GABAergic terminals. |
HSF1 transcription factor manipulation; electrophysiology (mIPSC recording); RT-PCR; pharmacological HSF1 inhibition |
Frontiers in integrative neuroscience |
Medium |
24376402
|
| 2014 |
VAMP2 (but not VAMP3) selectively mediates cAMP-stimulated NKCC2 exocytic delivery and surface expression in renal thick ascending limb cells. NKCC2 co-immunoprecipitates with VAMP2, and PKA activation enhanced VAMP2-NKCC2 co-immunoprecipitation. In vivo silencing of VAMP2 blocked cAMP-stimulated NKCC2 exocytic delivery. |
Co-immunoprecipitation; in vivo VAMP2/3 siRNA silencing; surface NKCC2 expression assay; VAMP2 exocytosis assay in renal cells |
The Journal of biological chemistry |
High |
25008321
|
| 2015 |
AP180, the VAMP2-specific endocytic adaptor, maintains the large pool of vesicular VAMP2 (70 copies/SV) that is required for efficient neurotransmission and SV reformation. Loss of AP180 causes moderate activity-dependent reduction of vesicular VAMP2 levels, defects in SV reformation, excitatory/inhibitory imbalance, epileptic seizures, and premature death. Further reduction in AP180-/-/Syb2+/- mice causes perinatal lethality. |
AP180 knockout mice; genetic interaction (AP180-/-/Syb2+/- double mutant); electrophysiology; electron microscopy of SV reformation |
Neuron |
High |
26412491
|
| 2017 |
The central small amino acids G100 and C103 in the VAMP2 transmembrane domain are critical for exocytosis by providing structural flexibility (alpha-helix/beta-sheet transitions) necessary for fusion pore opening and expansion. G100V/C103V mutation nearly abolished depolarization-evoked exocytosis, retarded initial fusion pore opening, hindered expansion, and led to premature pore closure. |
Site-directed mutagenesis (G100V/C103V); infrared spectroscopy (IRRAS, evanescent wave, ellipsometry); membrane capacitance; TIRF microscopy; ATP release measurement |
Scientific reports |
High |
28588281
|
| 2019 |
Heterozygous de novo mutations in VAMP2 (two single-amino-acid deletions and three missense variants in the C-terminal SNARE motif) cause a neurodevelopmental disorder with hypotonia, intellectual disability, and autistic features. Reconstituted lipid-mixing fusion assay demonstrated impaired vesicle fusion for these variants. |
Human genetics (de novo mutation identification); reconstituted lipid-mixing fusion assay |
American journal of human genetics |
Medium |
30929742
|
| 2020 |
Different regions of the synaptic vesicle membrane regulate VAMP2 conformation for SNARE assembly. In-cell NMR spectroscopy showed dynamic membrane association of VAMP2 SNARE motif in mammalian cells; cholesterol-rich lipid raft regions markedly weaken membrane association of the VAMP2 SNARE motif, releasing it to facilitate SNARE complex assembly, whereas non-raft regions maintain stronger membrane association. |
In-cell NMR spectroscopy; mass-spectrometry-based lipidomic profiling; lipid raft isolation |
Nature communications |
High |
32210233
|
| 2020 |
The complexin accessory helix suppresses spontaneous exocytosis by capturing the membrane-proximal C-terminal ends of both SNAP25 and VAMP2 prior to fusion in a reconstituted fusion assay. Site- and stage-specific photo-cross-linking revealed direct binding, and corresponding complexin interface mutants selectively increased spontaneous neurotransmitter release in living neurons. |
Reconstituted fusion assay; site-specific photo-cross-linking; neuronal electrophysiology |
Cell reports |
High |
32698012
|
| 2020 |
Munc18-1 helices 11 and 12 (domain 3a) interact with the VAMP2 SNARE motif covering layers -4 to +5; residue Q301 plays a pivotal role. A VAMP2-binding-deficient Munc18-1 Q301D mutant does not stimulate lipid mixing in reconstituted fusion and severely reduces synaptic transmission in Munc18-1-deficient neurons, demonstrating that Munc18-1/VAMP2 interaction is essential for SNARE templating. |
Comparative structure modeling; site-specific photo-cross-linking with unnatural amino acid Bpa; reconstituted vesicle docking/fusion assay; neuronal electrophysiology in Munc18-1-deficient neurons |
eNeuro |
High |
33055194
|
| 2020 |
Tetanus-insensitive (TI)-VAMP2 restores synaptic vesicle (SV) fusion in TeNT-treated hippocampal neurons but does not rescue dense core vesicle (DCV) fusion, despite TI-VAMP2 being targeted to and co-transported with DCVs. This demonstrates that VAMP2 is sufficient for SV fusion but that DCV fusion requires a distinct, unknown SNARE mechanism. |
Tetanus neurotoxin light chain expression; TI-VAMP2 rescue; live-cell imaging of DCV and SV fusion; VAMP1-null mutant analysis |
Scientific reports |
High |
32616842
|
| 2021 |
VAMP2 mediates most exocytosis from recycling endosomes (RE) in neuronal dendrites for LTP expression. However, VAMP4 mediates the majority of dendritic RE exocytosis, while VAMP2 plays only a minor role in RE exocytosis. LTP induction increases exocytosis from both VAMP2- and VAMP4-labeled organelles; VAMP4 knockdown decreases TfR recycling but increases AMPAR recycling, demonstrating VAMP2 and VAMP4 sort AMPARs and TfRs into separate endosomal populations. |
VAMP2/4 knockdown; TIRF live imaging; LTP electrophysiology; tetanus toxin-mediated VAMP2 cleavage |
Cell reports |
High |
34496238
|
| 2022 |
VAMP2/3-mediated exocytosis drives membrane expansion within myelin sheaths to initiate wrapping and power sheath elongation in oligodendrocytes. Genetic inactivation of VAMP2/3 in myelinating oligodendrocytes caused severe hypomyelination and premature death. Mass spectrometry revealed that VAMP2/3 incorporates axon-myelin adhesion proteins at the oligodendrocyte surface that are collectively required to form nodes of Ranvier. |
Conditional genetic knockout of VAMP2/3 in oligodendrocytes; live imaging; mass spectrometry of surface proteins |
Nature communications |
High |
36151203
|
| 2024 |
VAMP2 orchestrates alpha-synuclein (alphaSYN) phase separation both in vitro and in cells through electrostatic interactions between the VAMP2 juxtamembrane domain and the alphaSYN C-terminal region. VAMP2 binding induces co-condensate formation that sequesters vesicles and attracts complexin-1 and -2, and protects alphaSYN against forming aggregation-prone oligomers and fibrils. |
In vitro phase separation assays; live-cell condensate imaging; mutagenesis of charged residues; atomic force microscopy; fluorescence microscopy |
Nature cell biology |
High |
38951706 38951707
|
| 2024 |
The juxtamembrane region of VAMP2 directly interacts with the C-terminal region of alpha-synuclein through charged residues to regulate alphaSYN's function in clustering SVs and promoting SNARE complex assembly by inducing a multi-component condensed phase. VAMP2 binding protects alphaSYN against forming aggregation-prone oligomers and fibrils in condensates. |
Structural analysis; in vitro condensate formation; mutagenesis of charged residues; SV clustering assay; SNARE assembly assay; aggregation kinetics |
Nature cell biology |
High |
38951706
|
| 2024 |
VAMP2 is required for keratinocyte enucleation and epidermal differentiation through regulation of nucleophagy. VAMP2 binds FIP200, an autophagy protein, and both are required for enucleation. Deletion of VAMP2 leads to aberrant skin stratification and enhances cutaneous carcinogenesis in vivo. |
Genome-wide shRNA screen; in vivo VAMP2 deletion; quantitative proteomics; co-immunoprecipitation with FIP200 |
Developmental cell |
High |
38810653
|
| 2012 |
VAMP-2 is the v-SNARE involved in regulated surfactant secretion from alveolar type II cells. VAMP-2 and VAMP-8 are enriched in lamellar body fractions, but only the cytoplasmic domain of VAMP-2 (not VAMP-8) inhibited surfactant secretion in type II cells, and VAMP-2 co-localizes with the lamellar body marker protein LB-180. |
Subcellular fractionation; immunochemistry co-localization; cytoplasmic domain inhibition assay in type II cells |
Cell biology international |
Medium |
22571236
|
| 2010 |
VAMP-1 and VAMP-2, but not VAMP-3, regulate ANP release from atrial cardiac myocytes. VAMP-1 and VAMP-2 co-sediment and co-localize with ANP, form a SNARE complex with syntaxin-4 inside cardiac myocytes, and knockdown of VAMP-1 or VAMP-2 (but not VAMP-3) blocks regulated ANP release. |
Co-sedimentation; co-localization; co-immunoprecipitation of SNARE complex; siRNA knockdown; ANP secretion assay |
Journal of molecular and cellular cardiology |
Medium |
20801128
|
| 2010 |
Complexin 2 interacts with VAMP2 (as well as syntaxins 3 and 4) in pancreatic acini and regulates zymogen granule exocytosis. Introduction of recombinant complexin 2 inhibited Ca2+-stimulated secretion up to ~50%, and mutations of the central alpha-helical domain reduced SNARE binding and abolished inhibitory activity. An R59H mutation did not alter SNARE binding but augmented Ca2+-stimulated secretion by 130%. |
Co-immunoprecipitation; permeabilized acini reconstitution assay; site-directed mutagenesis; immunofluorescence co-localization |
The Journal of biological chemistry |
High |
20829354
|
| 2015 |
VAMP2 co-localizes with and mediates fusion of recycling vesicles containing transferrin receptor at the plasma membrane. VAMP2 depletion in HeLa cells suppressed recycling vesicle exocytosis, establishing a non-neuronal role for VAMP2 in transferrin receptor recycling downstream of the exocyst complex. |
siRNA depletion; co-localization microscopy; recycling vesicle exocytosis assay in HeLa cells |
Biology open |
Medium |
26092867
|
| 2020 |
Amyloid-beta peptides (Abeta40 and Abeta42) co-localize with VAMP2 in neuronal cells and decrease interactions between the N-termini of VAMP-2 and SNAP-25, disrupting SNARE complex formation critical for synaptic vesicle docking and fusion. |
FRET/FLIM imaging of Cer-VAMP2 and Cit-SNAP25 in N2A cells; confocal microscopy |
Journal of Alzheimer's disease : JAD |
Medium |
32675412
|
| 1997 |
VAMP-2 is present on gastric parietal cell tubulovesicles containing H+/K+-ATPase together with Rab11, Rab25, and SCAMPs, as demonstrated by immunoisolation on magnetic beads. The presence of VAMP-2 on immunoisolated H+/K+-ATPase-containing tubulovesicles supports their role in regulated vesicle fusion during parietal cell secretion. |
Gradient fractionation; immunoisolation on magnetic beads; Western blot analysis |
The Biochemical journal |
Medium |
9230141
|
| 1999 |
An alternatively spliced isoform of rat VAMP-2, called VAMP-2B (retaining an intron, altering the carboxy-terminal end), co-localizes with endogenous VAMP-2 in PC12 cells after subcellular fractionation, indicating that alternative splicing at the C-terminus does not affect VAMP-2 sorting. |
cDNA cloning; myc-epitope tagging; subcellular fractionation in PC12 cells |
FEBS letters |
Low |
10371166
|
| 2015 |
VAMP2-NRG1 is an oncogenic fusion gene that results from interchromosomal translocation; the fusion protein is membrane-bound, displays the NRG1 EGF-like domain extracellularly, and activates ERBB2/ERBB3 phosphorylation and downstream AKT/ERK signaling to promote anchorage-independent growth of lung adenocarcinoma cells. |
Whole-transcriptome sequencing; RT-PCR; immunoblotting for ERBB2/3/AKT/ERK phosphorylation; soft agar colony assay |
Journal of thoracic oncology |
Medium |
26134228
|
| 2023 |
VAMP2 cleavage in oligodendrocyte lineage cells impairs the maturation of premyelinating oligodendrocytes into functional myelinating cells in the developing mouse spinal cord. VAMP2/3-cleaved OLs stall at the premyelinating stage and show elevated Fyn kinase expression, indicating that VAMP2/3 activity is required for the premyelinating-to-myelinating transition. |
In vivo toxin-mediated VAMP2/3 cleavage in OL lineage; immunohistochemistry for Fyn; electron microscopy of myelin |
The Journal of neuroscience |
Medium |
37620160
|