Information on EC 3.6.5.2 - small monomeric GTPase and Organism(s) Homo sapiens

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EC NUMBER
COMMENTARY hide
3.6.5.2
-
RECOMMENDED NAME
GeneOntology No.
small monomeric GTPase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GTP + H2O = GDP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphorous acid anhydride hydrolysis
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
GTP phosphohydrolase (cell-regulating)
A family of about 50 enzymes with a molecular mass of 21 kDa that are distantly related to the alpha-subunit of heterotrimeric G-protein GTPase (EC 3.6.5.1). They are involved in cell-growth regulation (Ras subfamily), membrane vesicle traffic and uncoating (Rab and ARF subfamilies), nuclear protein import (Ran subfamily) and organization of the cytoskeleton (Rho and Rac subfamilies).
CAS REGISTRY NUMBER
COMMENTARY hide
9059-32-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
GTP + H2O
GDP + phosphate
show the reaction diagram
guanosine 5'-O-(3-thio)triphosphate + H2O
guanosine 5'-diphosphate + thiophosphate
show the reaction diagram
-
-
-
-
?
guanosine 5'-O-(3-thiotriphosphate) + H2O
guanosine 5'-O-diphosphate + thiophosphate
show the reaction diagram
MEK kinase 1 + H2O
?
show the reaction diagram
-
RhoA binds and activates MEK kinase 1
-
-
?
p21-activated kinase 1 + H2O
activated p21-activated kinase 1 + ?
show the reaction diagram
-
Gamide signals Rac/Cdc42 to activate p21-activated kinase 1
-
-
?
Rho-activated kinase + H2O
activated Rho-activated kinase + ?
show the reaction diagram
-
Ggly signals Rac/Cdc42 to activate Rho-activated kinase
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
GTP + H2O
GDP + phosphate
show the reaction diagram
p21-activated kinase 1 + H2O
activated p21-activated kinase 1 + ?
show the reaction diagram
-
Gamide signals Rac/Cdc42 to activate p21-activated kinase 1
-
-
?
Rho-activated kinase + H2O
activated Rho-activated kinase + ?
show the reaction diagram
-
Ggly signals Rac/Cdc42 to activate Rho-activated kinase
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2E)-2-[(2,5-dimethyl-1-phenyl-1H-pyrrol-3-yl)methylidene][1,3]thiazolo[3,2-a]benzimidazol-3(2H)-one
inhibition of GEF-Rac1 interaction (selective for Trio)
2-(morpholin-4-ylmethyl)-5-[(5-[[7-(trifluoromethyl)quinolin-4-yl]sulfanyl]pentyl)oxy]-4H-pyran-4-one
inhibition of Rac1 nucleotide binding possiblly using an allosteric mechanism
2-amino-8-hydroxy-9-[3-hydroxy-2-(hydroxymethyl)cyclopentyl]-5,9-dihydro-6H-purin-6-one
inhibition of Rac1-dependent NADPH oxidase activity
3-(2-hydroxyphenyl)-N-[4-(piperidin-1-ylsulfonyl)phenyl]-1H-pyrazole-5-carboxamide
inhibition of GEF-Rac1 interaction (Tiam1, Trio, and Vav2), the compound inhibits lamellipodia formation and smooth muscle cell migration
5'-p-fluorosulfonylbenzoylguanosine
-
irreversible substrate analogue-binding
9-methoxy-5-(3-nitrophenyl)-2-phenyl-3,10b-dihydropyrazolo[1,5-c][1,3]benzoxazine
inhibition of effector-Rac1 interaction (p67phox)
atorvastatin
-
marketed as Lipitor, i.e. [R-(R*,R*)]-2-(4-fluorophenyl)-beta,delta-dihydroxy-5-(1-methylethyl)-3-phenyl-4-[(phenylamino)carbonyl]-1H-pyrrole-1-heptanoic acid, inhibits RhoA activity by reducing Rho geranylgeranylation
Calmodulin
-
binding of calmodulin to GST-immobilized Kir/Gem peptide inhibits GTP binding
CCG-1423
-
-
CCG-977
-
-
cerivastatin
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marketed as Baycol/Lipobay, i.e. (E,3R,5S)-7-[4-(4-fluorophenyl)-5-(methoxymethyl)-2,6-dipropan-2-yl-pyridin-3-yl]-3,5-dihydroxy-hept-6-enoic acid, RhoA inhibitor
Clostridium botulinum exoenzyme C3
-
specific inhibitor
-
EDTA
inhibits nucleotide binding to Ras; inhibits nucleotide binding to Ras; inhibits nucleotide binding to Ras
GTP
-
inhibition of GTP[S]-binding at increasing concentrations
GTPgammaS
-
Arl6 is competitively inhibited by the increasing concentrations of non-radioactive GTPgammaS
guanine nucleotide dissociation inhibitor GDI
-
down-regulating GTPase activity, inhibition of nucleotide dissociation
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lovastatin
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marketed as Mevacor, i.e. 8-[2-(4-hydroxy-6-oxo-oxan-2-yl)ethyl]-3,7-dimethyl-1,2,3,7,8,8a-[hexahydronaphthalen-1-yl]2-methylbutanoate
N-[4-methoxy-3-(piperidin-1-ylsulfonyl)phenyl]-1H-indazole-3-carboxamide
inhibition of GEF-Rac1 interaction (Tiam1)
N4-(9-ethyl-9H-carbazol-3-yl)-N2-[3-(morpholin-4-yl)propyl]pyrimidine-2,4-diamine
inhibition of GEF-Rac1 interaction (selective for Vav2)
N6-(2-[[5-(diethylamino)pentan-2-yl]amino]-6-methylpyrimidin-4-yl)-2-methylquinoline-4,6-diamine
a selective inhibitor for Rac1. The small molecule fits into the surface groove of Rac1 involved in the binding with GEFs, thus interfering with the Tiam1-Rac1 interaction
NSC23766
-
not very potent inhibitor of Rac1 and Cdc42
p21 activated kinase
PAK, inhibits nucleotide dissociation from enzyme; PAK, inhibits nucleotide dissociation from enzyme
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Rap1Gap
-
the GTPase activating protein catalyzes the hydrolysis of GTP by its asparagine side chain rendering Rap1 inactive
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Rap1GAP1
-
a GTPase-activating protein that inhibits Rap1 activity
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simvastatin
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marketed as Zocor, i.e. [(1S,3R,7R,8S,8aR)-8-[2-[(2R,4R)-4-hydroxy-6-oxo-oxan-2-yl]ethyl]3.7-dimethyl]-1,2,3,7,8,8a-[hexahydronaphthalen-1-yl]2,2-dimethylbutanoate
statin
improves redox state in saphenous vein grafts in patients undergoing to coronary artery bypass grafting by inhibiting Rac1-mediated activation of NADPH oxidase
Yersinia outer protein T
-
Yersinia outer protein T is a cysteine protease that cleaves Rho protein directly upstream of the post-translationally modified cysteine, thereby releasing the GTPase from the membrane leading to inactivation, farnesylated RhoA is a preferred substrate of Yersinia outer protein T compared with the geranylgeranylated GTPase, geranylgeranylated RhoA, however, is the preferred substrate for Yersinia outer protein T-catalyzed cleavage with a 3fold faster turnover rate over Rac and Cdc42
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alphabeta-tubulin
soluble alphabeta-tubulin acts as a constitutively active Rheb activator that performs direct Rheb-binding, the deacetylated form has a high binding affinity for Rheb. Deacetylated soluble tubulin has a positive role in Rheb function by increasing the GTP-bound Rheb levels. Overexpression of alpha-tubulin K40A increases Rheb-induced S6K1 phosphorylation
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angiotensin II
stimulates Rac1 as cardiovascular stimulus
C3G guanine nucleotide exchange factor
-
-
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CalDAG-GEF
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a guanine nucleotide exchange factor
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cAMP
-
-
cytotoxic necrotizing factor 1
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activation of Rho proteins by the cytotoxic necrotizing factor 1 (CNF1) from Escherichia coli, while the isomeric cytotoxicnecrotizing factor from Yersinia pseudotuberculosis (CNFy) drives GTP-loading of basal RhoB but fails tocause activation of the rhoB promoter and thus its expression. CNF1 inhibits cytokinesis and induces the formation of bi-nucleated (tetraploid) cells. Cytotoxic-necrotizing factors encompass a class of auto-transporter toxins produced by Escherichia coli (CNF1-3) or Yersiniapseudotuberculosis (CNFy). CNF1 deamidates Gln63 in RhoA. CNF1-induced RhoB response depends on the deamidation of Rho proteins
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diacylglycerol
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Ect2cat
RhoA activation
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endothelin-1
stimulates Rac1 as cardiovascular stimulus
Epidermal growth factor
stimulates Rac1 as cardiovascular stimulus
farnesylthiosalicylic acid
-
markedly enhances RhoA level and activity
forskolin
-
the adenylyl cyclase activator strongly and specifically activates Rap1 in microvascular smooth muscle cells
GTP
the activation state of Rac1 depends on the release of guanosine diphosphate and the binding of guanosine triphosphate. This cycling is regulated by the guanine nucleotide exchange factors, as activators, and by the GTPase activating proteins; the activation state of Rac1 depends on the release of guanosine diphosphate and the binding of guanosine triphosphate. This cycling is regulated by the guanine nucleotide exchange factors, as activators, and by the GTPase activating proteins; the activation state of Rac1 depends on the release of guanosine diphosphate and the binding of guanosine triphosphate. This cycling is regulated by the guanine nucleotide exchange factors, as activators, and by the GTPase activating proteins. The activation process of Rac1 combines the GDP/GTP switch, catalyzed by GEF and GAP, with a cytosol/membrane alternation, regulated by GDI and protein prenylation, detailed overview
GTPase activating protein GAP
-
guanine nucleotide exchange factor Epac
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specifically increases Rap1 activity
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guanine nucleotide exchange factor GEF
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guanine nucleotide exchange factors
GEF, GTP-binding to small GTPases is catalyzed by GEFs, Trp71 of Rac1 is a critical site for the discrimination of a subset of GEF, including Tiam1 and Trio
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heparin-binding epidermal growth factor-like growth factor
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induces rapid and strong RhoA activation
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Insulin
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activation of Rac, assembly of actin filaments
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Insulin-like growth factor
stimulates Rac1 as cardiovascular stimulus
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interleukin 1beta
stimulates Rac1 as cardiovascular stimulus
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lysophosphatidic acid
melanophilin
-
Rab27a effector
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p21-activated kinase 1
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interacts with Rac and Cdc42
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PDZ-GEF1
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a guanine nucleotide exchange factor
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PDZ-GEF2
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a guanine nucleotide exchange factor
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platelet derived growth factor
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Rac activation, assembly of actin filaments
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platelet-derived growth factor
stimulates Rac1 as cardiovascular stimulus
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PLCepsilon
-
a guanine nucleotide exchange factor
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Prorenin
stimulates Rac1 as cardiovascular stimulus
-
Rac-specific GEF Tiam1
activates Rac3, but less efficiently than the Rac isoform Rac2; stimulates the GDP exchange reaction, increases the GDP dissociation rate, activates Rac1 less efficiently than Rac2; stimulates the GDP exchange reaction, increases the GDP dissociation rate, activates Rac2 more efficiently than the Rac isoforms Rac1 and Rac3
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Ras
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cross-talk between Ras- and Rho subfamilies, activation of Rac
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RasGRP2
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a guanine nucleotide exchange factor
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SOS980-989 peptide
the recombinant SOS980-989 peptide contains the H-Ras-SOScat interaction contact region, which is known to be able to compete with SOScat for binding to H-Ras; the recombinant SOS980-989 peptide contains the H-Ras-SOScat interaction contact region, which is known to be able to compete with SOScat for binding to H-Ras
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SOScat
activation of Ras; activation of Ras
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sphingosine 1 phosphate
stimulates Rac1 as cardiovascular stimulus
T-lymphoma invasion and metastasis factor 1
Tiam1, a GEF, structure analysis in complex with Rac1, specific site of GEF-Rac1 interaction, in particular Trp71
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thrombin
stimulates Rac1 as cardiovascular stimulus
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tumor necrosis factor-alpha
stimulates Rac1 as cardiovascular stimulus
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U46619
-
small GTPase RhoA is activated by the activation of functional receptors for thromboxane A2 like U46619
vascular endothelial growth factor
stimulates Rac1 as cardiovascular stimulus
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additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0005
GTP
-
quite low, increased GTP hydrolysis by action of guanine nucleotide release proteins, promoting replacement of bound GDP by GTP
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001
GDP
-
inhibition of binding of GTP[S]
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
(2E)-2-[(2,5-dimethyl-1-phenyl-1H-pyrrol-3-yl)methylidene][1,3]thiazolo[3,2-a]benzimidazol-3(2H)-one
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.005
2-(morpholin-4-ylmethyl)-5-[(5-[[7-(trifluoromethyl)quinolin-4-yl]sulfanyl]pentyl)oxy]-4H-pyran-4-one
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.156
2-amino-8-hydroxy-9-[3-hydroxy-2-(hydroxymethyl)cyclopentyl]-5,9-dihydro-6H-purin-6-one
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.0087 - 0.0088
3-(2-hydroxyphenyl)-N-[4-(piperidin-1-ylsulfonyl)phenyl]-1H-pyrazole-5-carboxamide
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.01
9-methoxy-5-(3-nitrophenyl)-2-phenyl-3,10b-dihydropyrazolo[1,5-c][1,3]benzoxazine
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.024
N-[4-methoxy-3-(piperidin-1-ylsulfonyl)phenyl]-1H-indazole-3-carboxamide
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.0011
N4-(9-ethyl-9H-carbazol-3-yl)-N2-[3-(morpholin-4-yl)propyl]pyrimidine-2,4-diamine
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
0.05
N6-(2-[[5-(diethylamino)pentan-2-yl]amino]-6-methylpyrimidin-4-yl)-2-methylquinoline-4,6-diamine
Homo sapiens;
P15153, P60763, P63000
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.001
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
assay at
7.6
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
assay at room temperature; assay at room temperature; assay at room temperature
25
assay at; assay at
30
-
assay at
37
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
human alveolar epithelial cell HAEC exposed to hypoxia, RhoA/ROCK activation is necessary for Na,K-ATPase endocytosis via a mechanism that requires mitochondrial reactive oxygen species
Manually annotated by BRENDA team
two isoforms of RASL11A transcript with different sizes of 1.6 and 1.2 kb
Manually annotated by BRENDA team
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mRNA levels of RasL10B are down-regulated in all breast cancer cells tested (HBL100, MCF7, MDA-MB-468, MDA-MB-231 and MDA-MB-435. RasL10B) is a member of ras superfamily with tumor suppressor potential
Manually annotated by BRENDA team
two isoforms of RASL11A transcript with different sizes of 1.6 and 1.2 kb
Manually annotated by BRENDA team
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Ras is detected in glomerular cells, proximal convoluted tubule cells, distal convoluted tubule cells, cortical collecting tubule cells, medullar collecting duct cells, interstitial cells and subcapsular fibroblast. Ras plays a role in renal fibrosis
Manually annotated by BRENDA team
lung cancer patients with metastasis and poor survival show low hRAB37 protein expression coinciding with low TIMP1 in tumours, hRAB37 is downregulated mainly by promoter hypermethylation in lung cancer cells and patients, low mRNA expression of hRAB37 gene is associated with tumour metastasis in lung cancer patients
Manually annotated by BRENDA team
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B-EBV lymphoblastoid cell line
Manually annotated by BRENDA team
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expression of Rap2C may be down-regulated diring the late phase of megakaryocyte differentiation and platelet formation
Manually annotated by BRENDA team
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Rho-mediated signaling in osteoblasts seems to introduce major biases to bone resorption
Manually annotated by BRENDA team
very weak expression of RASL11A
Manually annotated by BRENDA team
adjacent, two isoforms of RASL11A transcript with different sizes of 1.6 and 1.2 kb, the abundance of the RASL11A transcript is diminished in prostate tumors in comparison to normal adjacent prostate tissue
Manually annotated by BRENDA team
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small Rho GTPases are important for acinus formation
Manually annotated by BRENDA team
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proliferation of dermal fibroblasts is crucial for the maintenance of skin. Rac1 activates proliferation of normal fibroblasts through stimulation of c-myc phosphorylation without affecting ERK1/2 activity
Manually annotated by BRENDA team
RhoH is expressed strongest in T-lymphocytes and acts as a positive regulatory factor for thymocyte selection and T-cell receptor signalling
Manually annotated by BRENDA team
; two isoforms of RASL11A transcript with different sizes of 1.6 and 1.2 kb
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
Rab21 is predominantly localized to the early endocytic pathway, on vesicles containing early-endosomal antigen 1, transferrin receptor and internalized ligands
Manually annotated by BRENDA team
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Rab1b-labeled vesicles, distribute peripherally from the Golgi perinuclear area
-
Manually annotated by BRENDA team
part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities
Manually annotated by BRENDA team
-
human Mammarian enabled protein colocalizes with Rac1 in lamellipodia
Manually annotated by BRENDA team
part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities; part of invadosomes, specialized plasma-membrane actin-based microdomains that combine adhesive properties with matrix degrading and/or mechanosensor activities
Manually annotated by BRENDA team
colocalization with tissue inhibitor of metalloproteinase 1 (TIMP1)
-
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
UNIPROT
ORGANISM
Homo sapiens;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22000
-
SDS-PAGE
23000
-
complex of 1 * 23000 for Cdc42Hs + 1 * 26000 for regulatory protein LyGDL, SDS-PAGE, immunostaining
26000
-
complex of 1 * 23000 for Cdc42Hs + 1 * 26000 for regulatory protein LyGDL, SDS-PAGE, immunostaining
51000
x * 51000, GFP-Rab21, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 51000, GFP-Rab21, SDS-PAGE
dimer
-
complex of 1 * 23000 for Cdc42Hs + 1 * 26000 for regulatory protein LyGDL, SDS-PAGE, immunostaining
monomer
-
1 * 22000, SDS-PAGE
additional information
structural core of RhoGTPases: the G domain, the Rho insert region, and the C-terminus of RhoGTPases, structure analysis, detailed overview. The hypervariable region also functions as protein binding site and determines, at least in part, specific binding to regulatory or effector proteins. In most RhoGTPases, including RhoA and RhoC, this region of around 10 residues harbors a polybasic stretch that binds the inner leaflet of the plasma membrane. RhoA, RhoB and RhoC have a very similar structure of the G domain and the insert region; structural core of RhoGTPases: the G domain, the Rho insert region, and the C-terminus of RhoGTPases, structure analysis, detailed overview. The hypervariable region also functions as protein binding site and determines, at least in part, specific binding to regulatory or effector proteins. In most RhoGTPases, including RhoA and RhoC, this region of around 10 residues harbors a polybasic stretch that binds the inner leaflet of the plasma membrane. RhoA, RhoB and RhoC have a very similar structure of the G domain and the insert region; structural core of RhoGTPases: the G domain, the Rho insert region, and the C-terminus of RhoGTPases, structure analysis, detailed overview. The hypervariable region also functions as protein binding site and determines, at least in part, specific binding to regulatory or effector proteins. In most RhoGTPases, including RhoA and RhoC, this region of around 10 residues harbors a polybasic stretch that binds the inner leaflet of the plasma membrane. RhoA, RhoB and RhoC have a very similar structure of the G domain and the insert region
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
lipoprotein
phosphoprotein
PKA- and PKG-mediated phosphorylation of RhoA on Ser188, which is located in the hypervariable C-terminus. This phosphorylation serves as a Rho-inactivating signal, as it promotes GDI binding and membrane dissociation and protects RhoA from proteolytic degradation. In addition, this phosphorylation interferes with RhoA binding to ROCK further underscoring a role of the C-terminus in effector interactions; RhoC, but not RhoA, is phosphorylated on Ser73 in the alpha2-helix by the kinase Akt in SUM149 breast cancer cells54 although Ser73 is highly conserved among all 3 Rho-isoforms. Intriguingly, phosphorylation of RhoC is required for downstream signaling and invasiveness, which contrasts markedly with the inactivating phosphorylation at Ser188 in RhoA
side-chain modification
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1.65 A resolution, complex of RhoA-rhoGAP with the transition-state analogue GDPAIF4-, hanging drop vapour diffusion technique
-
Arf1 complexed to GDP, common G domain topology; Rac1, active conformation
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complex of Cdc42Hs-GMPP-NP-RhoGAP
-
crystals are obtained at 20°C by vapour diffusion using a crystallization robot. The crystals are found to belong to space group P2(1), with unit-cell parameters a = 52.2, b = 58.6, c =53.4 A, beta= 97.9°, and contains two Rad molecules in the crystallographic asymmetric unit, 1.8 A resolution
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GTPase domain of human Rheb recombinantly expressed in Escherichia coli, purified and cocrystallized in complexes with GDP, GTP and GppNHp using the hanging-drop vapour-diffusion method. Crystals of the hRheb/GDP complex belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 44.5, b = 52.3, c = 70.6 A. The hRheb/GppNHp complex crystallizes in two crystal forms: one has the same space group and unit-cell parameters as the hRheb/GDP complex and the other belongs to space group C222(1), with unit-cell parameters a = 102.9, b = 99.2, c = 48.0 A. The hRheb/GTP complex also crystallizes in two crystal forms: one belongs to space group C222(1), with unit-cell parameters a = 102.4, b = 98.3, c = 47.9 A, and the other belongs to space group P2(1), with unit-cell parameters a = 77.3, b = 47.9, c = 71.9 A, beta = 89°. All these crystals diffract X-rays to better than 2.8 A resolution
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hanging-drop method, Rad in the GDP-bound form at 1.8 A resolution
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hanging-drop vapor-diffusion method, Rab11b-GDP and Rab11b-GppNHp crystal structures solved to 1.55 and 1.95 A resolution, respectively
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solution structure of free RalB bound to the GTP analogue GMPPNP, nuclear magnetic resonance spectroscopy
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vapor diffusion hanging-drop method, three-dimensional crystal structure of Rab4a in its GppNHp-bound state to 1.6 A resolution and in its GDP-bound state to 1.8 A resolution
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, little loss of activity, several weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
glutathione Sepharose 4B column chromatography
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glutathione-agarose bead chromatography and Superdex-75 gel filtration
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GST-tagged Rheb by glutathione affinity chromatography, recombinant enzyme from HEK-293 cells by tandem affinity purification through IgG-binding and anti-Flag affinity chromatography
GTPase core domain overexpressed in Escherichia coli
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GTPase domain of human Rheb recombinantly expressed in Escherichia coli
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Q Sepharose column chromatography, ammonium sulfate precipitation, glutathione 4B-Sepharose bead chromatography, and Superdex 200 gel filtration
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recombinant
-
recombinant enzymes, affinity chromatography
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recombinant glutathione S-transferase (GST)-Wrch- and hexa-histidine (His6)-tagged Wrch-1 protein from Escherichia coli
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recombinant Rac1; recombinant Rac2; recombinant Rac3
recombinant wild-type and mutant Rab27a
-
to homogeneity, chromatography techniques
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of rhoB mRNA level by semi-quantitative real-time RT-PCR
-
Cdc42, expression in Escherichia coli
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coexpression of GFP-LC3 and either Sar1 wild-type or the mutants Sar1 H79G or Sar1 T39N in CHO cells, effect of overexpression of Sar1 and its mutants in the secretory pathway, overview. Overexpression of GFP-tagged Rab1b wild-type or mutant Rab1b in CHO cells. Overexpression of GFP-tagged Rab1b N121I mutant in Vero monkey cells and human HaCaT cells
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complete coding sequence of ARL9, from testis, maps to chromosome 4q12; complete coding sequence of RASL11A, from prostate, maps to chromosome 13q12.2
expressed in Escherichia coli
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expressed in Escherichia coli BL21 cells
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21-Codon-Plus(DE3) cells, and in MDCKII and Flp-In-293 cells
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expressed in Escherichia coli strain BL21(DE3)
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expression in COS-7 cells
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expression in COS7 cells
expression in Escherichia coli
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expression in Escherichia coli; expression in Sf9 cells
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expression in Sf9 cells
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gene rad, recombinant expression of GST-tagged Rad in Escherichia coli
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GTPase core domain is overexpressed in Escherichia coli
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GTPase domain of human Rheb recombinantly expressed in Escherichia coli
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Rab1A, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs; Rab1B, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs; Rab2A, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs; Rab2B, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs; Rab6A, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs; Rab8B, genome-wide screening for human Rabs that are involved in Golgi morphology in HeLa-S3 cells using specific siRNAs
Rab21 cDNA from K-562 cell, expression in HeLa cells CCL-2 and in hepatocellular carcinoma Hep3B cells
Rac1, expression in Escherichia coli as glutathione S-transferase fusion protein; Rac2, expression in Escherichia coli as glutathione S-transferase fusion protein; Rac3, expression in Escherichia coli as glutathione S-transferase fusion protein
recombinant expression of dual tagged Rheb in HEK-293 cells or of GST-tagged Rheb
recombinant expression of GST-tagged RhoA wild-type (amino acids 1-193) in Escherichia coli; recombinant expression of H-Ras wild-type (amino acids 2-189) and H-Ras mutant Q61G (amino acids 2-189) in Escherichia coli; recombinant expression of K-Ras wild-type (amino acids 2-189) in Escherichia coli
wild-type and mutant Rab27a cDNA, expression in Escherichia coli BL21 and in 293T cells, expression of mutant Rab27a in the mouse melanocyte cell line melan-a
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
hRAB37 is downregulated mainly by promoter hypermethylation in lung cancer cells
knockdown of Rab isoform Rab2A induces fragmentation of the Golgi in HeLa-S3 cells, its phenotype is rescued by re-expression of its siRNA-resistant Rab construct alone, not by any of the other five Rab isozymes
overexpression of any type of Rhebs (wild-type, constitutively active form, and dominant-negative form) does not affect the intact microtubule structure, and the vesicle-formed Rheb does not co-localize with the microtubule network
RhoB is the only member of the Rho subfamily of small GTPases, which is classified as an immediate early gene product. RhoB is up-regulated in response to growth factors as well as cytotoxic and genotoxic agents. Clostridial glucosylating toxins evoke pronounced RhoB expression, based on the inactivation of Rho/Ras proteins. Long lasting expression of RhoB in cultured cells upon activation of Rho proteins by the cytotoxic necrotizing factor 1 (CNF1) from Escherichia coli. Deamidase-deficient CNF1-C866S fails to activate the rhoB promoter, CNF1-C866S only faintly induces expression of rhoB mRNA and RhoB protein. Critical role of Rac1 in CNF1-induced RhoB expression, overview
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A152P
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mutation dramatically affects both GTP and GDP nucleotide-binding activity of Rab27a, probably by disrupting protein folding
chimeras between RhoA/Cdc42
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residues 85-122 contain specific binding determinants for guanosine nucleotide activating protein, p190
D60I
site-directed mutagenesis
D60K
site-directed mutagenesis
D90A
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RhoA, significant decrease in p190-mediated stimulation of GTPase activity
G169A
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the mutation causes the Bardet-Biedl syndrome, the mutant protein is destabilized probably due to the weak affinity to guanine nucleotides
H79G
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a GTPase-defective Sar1 mutant. Overexpression of the GTPase-defective mutant Sar1 H79G, in its GTP-bound form, allows the development of ERES, but the transport to the Golgi apparatus is blocked because this mutant inhibits COPII coat dissociation, and its expression induces Golgi fragmentation as indicated by Rab6 dispersion and inhibits autophagosome formation
L130P
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mutation dramatically affects both GTP and GDP nucleotide-binding activity of Rab27a, probably by disrupting protein folding
L170W
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the mutation causes the Bardet-Biedl syndrome, the mutant protein is destabilized probably due to the weak affinity to guanine nucleotides
N-terminal truncation
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Kir/Gem peptide, C-terminal calmodulin-binding domain, high affinity for dansyl-calmodulin
N121I
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a dominant-negative GFP-tagged Rab1b mutant, inhibits autophagosome formation
Q67L
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a GTPase-deficient Rab1b mutant, the distribution of Rab1b in the acidic compartments responds to inhibition of autophagy
Q72L
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the mutant decreases the GTP hydrolysis activity and stabilizes the active form of the protein
Q89L
site-directed mutagenesis, a GTP-bound active mutant
S88D
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Cdc42, enhancement of GTP-hydrolysis
T31M
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the mutation causes the Bardet-Biedl syndrome, the mutant protein is destabilized probably due to the weak affinity to guanine nucleotides
T31N
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the guanine nucleotide-free Arl6 mutant protein is unstable and degraded in living cells
T31R
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the mutation selectively abrogates the GTP-binding ability of Arl6 without affecting GDP-binding/dissociating properties
T33N
mutant enzyme is defective in GTP binding, cells expressing mutant Rab21 show defects in endocytosis of transferrin and epidermal growth factor and fail to effectively deliver the latter ligand to late endosomes and lysosomes for degradation, subcellular distribution
T35A
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Rap1A, blocks abilitiy of Rap-GAP to stimulate GTP hydrolysis
T39N
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a dominant-negative Sar1 mutant, expression induces Golgi fragmentation as indicated by Rab6 dispersion and inhibits autophagosome formation
T43N
site-directed mutagenesis, a GDP-bound inactive mutant
W269G
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Kir/Gem peptide, abolish affinity for dansyl-calmodulin
W73G
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mutant protein has GTP-binding activity but is inefficient in hydrolyzing GTP, in contrast to Q78L it neither interacts with the Rab27a effector melanophilin nor modifies melanosome distribution and cytotoxic granule exocytosis, mutation may increase switch flexibility and thus impair the dynamics of the conformational changes associated with each nucleotide binding
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
nucleotide exchange kinetics with small GTPases can be used to obtain important information about the binding kinetics of GTP molecules under different activation states and with different activator molecules; nucleotide exchange kinetics with small GTPases can be used to obtain important information about the binding kinetics of GTP molecules under different activation states and with different activator molecules; nucleotide exchange kinetics with small GTPases can be used to obtain important information about the binding kinetics of GTP molecules under different activation states and with different activator molecules
medicine
pharmacology
the enzyme is a pharmacological target for the treatment of cardiovascular diseases; the enzyme is a pharmacological target for the treatment of cardiovascular diseases; the enzyme is a pharmacological target for the treatment of cardiovascular diseases