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Information on EC 3.6.4.6 - vesicle-fusing ATPase

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EC Tree
IUBMB Comments
A large family of ATP-hydrolysing enzymes involved in the heterotypic fusion of membrane vesicles with target membranes and the homotypic fusion of various membrane compartments. They belong to the AAA-type (_A_TPase _a_ssociated with a variety of cell _a_ctivities) ATPase superfamily. They include peroxin, which apparently is involved in Zellweger's syndrome.
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UNIPROT: P52917
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Word Map
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
Synonyms
n-ethylmaleimide-sensitive factor, cdc48, aaa-atpase, n-ethylmaleimide-sensitive fusion protein, atp6v0d2, n-ethylmaleimide sensitive factor, sec18p, sec18, cdc48p, pex1p, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
vacuolar protein sorting 4 protein
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N-ethylmaleimide sensitive fusion protein
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NEM-sensitive fusion protein
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SKD2 protein
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Vesicular-fusion protein NSF
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
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SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (vesicle-fusing)
A large family of ATP-hydrolysing enzymes involved in the heterotypic fusion of membrane vesicles with target membranes and the homotypic fusion of various membrane compartments. They belong to the AAA-type (_A_TPase _a_ssociated with a variety of cell _a_ctivities) ATPase superfamily. They include peroxin, which apparently is involved in Zellweger's syndrome.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
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?
additional information
?
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residues from helix 5 of the Vps2p subunit of ESCRT-III bind to the central pore of an asymmetric Vps4p hexamer in a manner that is dependent upon the presence of flexible nucleotide analogues that can mimic multiple states in the ATP hydrolysis cycle, quantitative binding studies. Vps2p helix 5 peptides bind the Vps4p ATPase cassette. Vps4 substrates are initially recruited by an MIM-MIT interaction that activates the Vps4 central pore to engage substrates and generate force, thereby triggering ESCRT-III disassembly. The Vps4p hexamer binds a single peptide
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
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-
?
additional information
?
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residues from helix 5 of the Vps2p subunit of ESCRT-III bind to the central pore of an asymmetric Vps4p hexamer in a manner that is dependent upon the presence of flexible nucleotide analogues that can mimic multiple states in the ATP hydrolysis cycle, quantitative binding studies. Vps2p helix 5 peptides bind the Vps4p ATPase cassette. Vps4 substrates are initially recruited by an MIM-MIT interaction that activates the Vps4 central pore to engage substrates and generate force, thereby triggering ESCRT-III disassembly. The Vps4p hexamer binds a single peptide
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
binding of substrates to the central pore of the Vps4 ATPase is autoinhibited by the microtubule interacting and trafficking (MIT) domain, substrate engagement is autoinhibited by the Vps4p-MIT-domain and the inhibition is relieved by binding of either Type 1 or Type 2 MIM elements, which bind the Vps4p MIT domain through different interfaces
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Vps2p helix 5 peptide C
stimulates Vps4p ATPase activity and stabilizes the hexamer
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additional information
binding of substrates to the central pore of the Vps4 ATPase is activated by microtubule interacting and trafficking interacting motifs (MIMs)
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme Vps4 belongs to the type I AAA ATPases
physiological function
the endosomal sorting complexes required for transport (ESCRT) pathway drives reverse topology membrane fission events within multiple cellular pathways, including cytokinesis, multivesicular body biogenesis, repair of the plasma membrane, nuclear membrane vesicle formation, and HIV budding. The AAA ATPase Vps4 is recruited to membrane necks shortly before fission, where it catalyzes disassembly of the ESCRT-III lattice. Vps4 substrates are initially recruited by an MIM-MIT interaction that activates the Vps4 central pore to engage substrates and generate force, thereby triggering ESCRT-III disassembly. Vps2p helix 5 peptides bind the Vps4p ATPase cassette. Two-step model for disassembly of the ESCRT-III complex by Vps4, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
VPS4_YEAST
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
437
0
48172
Swiss-Prot
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
Vps4p dimers form two distinct heptameric rings and accommodate AAA cassettes in a head-to-head fashion, X-ray crystallography
hexamer
eukaryotic Vps4 enzymes comprise the N-terminal MIT domain, an about 40-residue linker, a two-domain AAA ATPase cassette, a beta-domain that is inserted within the small domain of the ATPase cassette, and a C-terminal helix that binds against the large ATPase domain. Vps4 functions as a higher-order oligomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
sitting drop vapour diffusion method, at 19°C against a reservoir containing 0.1 M MES buffer (pH 5.9-6.5) and 0.8-1.9 M MgSO4
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E233Q
site-directed mutagenesis, an ATPase inactive mutant
E243A
site-directed mutagenesis, a pore loop mutant, displays essentially unchanged oligomerization, introducing E243A point mutation increases the affinity of peptide C binding by 3fold, shows reduced ATPase activity compared to the wild-type
E243A/E247A
site-directed mutagenesis, pore loop mutant, displays essentially unchanged oligomerization, the double mutant binds peptides 4fold (peptide C) or 10fold (peptide B) more tightly than wild-type Vps4p, shows reduced ATPase activity compared to the wild-type
E247A
site-directed mutagenesis, a pore loop mutant, displays essentially unchanged oligomerization, introducing E247A point mutation increases the affinity of peptide C binding by 4fold, shows reduced ATPase activity compared to the wild-type
Q216A
monomeric mutant form
R241A
site-directed mutagenesis, a pore loop 2 mutant, does not bind peptides with appreciable affinity
R251A
site-directed mutagenesis, a pore loop 2 adjacent mutant, does not bind peptides with appreciable affinity
T240A
site-directed mutagenesis, a pore loop mutant, displays essentially unchanged oligomerization
T240F
site-directed mutagenesis, a pore loop 2 mutant
T240K
site-directed mutagenesis, a pore loop 2 mutant
T240V
site-directed mutagenesis, a pore loop 2 mutant
W206A
site-directed mutagenesis, a pore loop 1 mutant, does not bind peptides with appreciable affinity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-chelating column chromatography and Superdex gel filtration
recombinant wild-type and mutant Vps4ps
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
recombinant expression of wild-type and mutant Vps4ps
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hartmann, C.; Chami, M.; Zachariae, U.; de Groot, B.L.; Engel, A.; Gruetter, M.G.
Vacuolar protein sorting: two different functional states of the AAA-ATPase Vps4p
J. Mol. Biol.
377
352-363
2008
Saccharomyces cerevisiae (P52917), Saccharomyces cerevisiae
Manually annotated by BRENDA team
Han, H.; Monroe, N.; Votteler, J.; Shakya, B.; Sundquist, W.I.; Hill, C.P.
Binding of substrates to the central pore of the Vps4 ATPase is autoinhibited by the microtubule interacting and trafficking (MIT) domain and activated by MIT interacting motifs (MIMs)
J. Biol. Chem.
290
13490-13499
2015
Saccharomyces cerevisiae (P52917)
Manually annotated by BRENDA team