Information on EC 2.4.1.B62 - small GTPase glucosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.B62
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
small GTPase glucosyltransferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-alpha-D-glucose + a small GTPase = UDP + D-glucosyl-[a small GTPase]
show the reaction diagram
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SYSTEMATIC NAME
IUBMB Comments
UDP-alpha-D-glucose:small GTPase glucosyltransferase
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain does not produce toxin A
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Manually annotated by BRENDA team
alpha-toxin; type A
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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cytotoxin A possesses an inherent cysteine protease activity, which is responsible for auto-cleavage of glucosylating toxins; cytotoxin B possesses an inherent cysteine protease activity, which is responsible for auto-cleavage of glucosylating toxins
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
UDP-alpha-D-glucose + Cdc42
UDP + D-glucosyl-Cdc42
show the reaction diagram
UDP-alpha-D-glucose + Cdc42Hs
UDP + D-glucosyl-Cdc42Hs
show the reaction diagram
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-
-
?
UDP-alpha-D-glucose + H-Ras
UDP + D-glucosyl-H-Ras
show the reaction diagram
UDP-alpha-D-glucose + K-Ras
UDP + D-glucosyl-K-Ras
show the reaction diagram
good substrate
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-
?
UDP-alpha-D-glucose + N-Ras
UDP + D-glucosyl-N-Ras
show the reaction diagram
good substrate
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-
?
UDP-alpha-D-glucose + Rac1
UDP + D-glucosyl-Rac1
show the reaction diagram
UDP-alpha-D-glucose + RalC
UDP + D-glucosyl-RalC
show the reaction diagram
good substrate
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-
?
UDP-alpha-D-glucose + Rap2A
UDP + D-glucosyl-Rap2A
show the reaction diagram
UDP-alpha-D-glucose + RhoA
UDP + D-glucosyl-RhoA
show the reaction diagram
UDP-alpha-D-glucose + RhoB
UDP + D-glucosyl-Rhob
show the reaction diagram
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-
-
?
UDP-alpha-D-glucose + RhoC
UDP + D-glucosyl-RhoC
show the reaction diagram
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-
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?
UDP-alpha-D-glucose + RhoG
UDP + D-glucosyl-RhoG
show the reaction diagram
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-
-
?
UDP-alpha-D-glucose + TC10
UDP + D-glucosyl-TC10
show the reaction diagram
good substrate
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-
?
UDP-alpha-D-glucose + TCL
UDP + D-glucosyl-TCL
show the reaction diagram
good substrate
-
-
?
UDP-N-acetyl-alpha-D-glucosamine + H-Ras
UDP + N-acetyl-alpha-D-glucosamine-H-Ras
show the reaction diagram
UDP-N-acetyl-alpha-D-glucosamine + RhoA
UDP + N-acetyl-D-glucosamine-RhoA
show the reaction diagram
additional information
?
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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
UDP-alpha-D-glucose + H-Ras
UDP + D-glucosyl-H-Ras
show the reaction diagram
UDP-alpha-D-glucose + Rap2A
UDP + D-glucosyl-Rap2A
show the reaction diagram
UDP-alpha-D-glucose + RhoA
UDP + D-glucosyl-RhoA
show the reaction diagram
additional information
?
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P16154
enzyme glucosylates and inactivates small GTPases of the Rho or Ras families, culminating in cytotoxicity
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
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stimulation, maximum activity at 1 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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2 mM, complete inhibition
additional information
toxin A undergoes autocatalytic processing. The glucosyltransferase activity of the TcdA(1-1832) protein is activated by autoproteolysis. The presence of the autoprotease domain, residues 543-800, is restricting the activity of the glucosyl transferase domain
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.004 - 0.018
UDP-alpha-D-glucose
0.026 - 0.96
UDP-N-acetyl-alpha-D-glucosamine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.39 - 0.59
UDP-alpha-D-glucose
0.12 - 0.66
UDP-N-acetyl-alpha-D-glucosamine
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
22.8 - 97.2
UDP-alpha-D-glucose
364
1.9 - 25.3
UDP-N-acetyl-alpha-D-glucosamine
1424
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43000
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x * 220000, x * 43000, SDS-PAGE
105000
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and 220000, PAGE of toxin B
220000
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and 105000, PAGE. The 220000 Da subunit of toxin B from strain 8864 migrates slightly faster than the subunit from strain 10463
307000
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x * 307000, SDS-PAGE
308000
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x * 308000, calculated
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
multimer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
catalytic fragment of toxin B in the presence of UDP-glucose and Mn2+, to 2.2 A resolution. Toxin B belongs to the glycosyltransferase type A family. The reaction proceeds probably along a single-displacement pathway. The C1'' donor carbon atom position is defined by the bound UDP and glucose. The relative orientation of toxin B and substrate RhoA is consistent with both being attached to a membrane
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crystal structures of the toxin A glucosyltransferase domain in the presence and absence of the co-substrate UDP-alpha-D-glucose, to 2.2 and 2.6 A resolution, respectively
enzyme contains an internal cysteine protease domain allosterically regulated by the eukaryotic-specific molecule inositol hexakisphosphate. Apo-cysteine protease is in dynamic equilibrium between active and inactive states. Inositol hexakisphosphate dramatically shifts this equilibrium towards an active conformer that is further restrained upon binding a suicide substrate. Residues within a beta-hairpin region functionally couple the inositol hexakisphosphate binding site to the active site
catalytic fragment, residues 1-551 to 2.85 A resolution. Geometry suggests that the reaction runs as a circular electron transfer in a six-membered ring, which involves the deprotonation of the nucleophile by the beta-phosphoryl group of the donor substrate UDP-glucose
catalytic fragment, residues 1-546, to 2.3 A resolution. Geometry suggests that the reaction runs as a circular electron transfer in a six-membered ring, which involves the deprotonation of the nucleophile by the beta-phosphoryl group of the donor substrate UDP-glucose
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purification of toxin B from strain 8864 that does not produce toxin A
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Bacillus megaterium
expression in Escherichia coli
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expression in Escherichia coli; expression in Escherichia coli
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expression of catalytic fragment, residues 1-546
expression of catalytic fragment, residues 1-551
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C698A
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mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
D587N
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mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
H653A
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mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
I383S
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mutation in toxin B, 67% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
I383S/Q385A
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mutation in toxin B, 23% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose. Mutation largely increases the acceptance of UDP-Nacetylglucosamine as a sugar donor for modification of RhoA
Q385A
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mutation in toxin B, 58% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
I383S
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mutation in toxin B, 67% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
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I383S/Q385A
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mutation in toxin B, 23% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose. Mutation largely increases the acceptance of UDP-Nacetylglucosamine as a sugar donor for modification of RhoA
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Q385A
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mutation in toxin B, 58% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
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S385/A387Q
F17K
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mutation strongly decreases binding to brain phosphatidylserine
I383S/Q385A
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mutation allow modification of Ras in the presence of UDP-N-acetyl-glucosamine and reduces the acceptance of UDP-glucose as a donor for glycosylation
K11I
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mutation moderately decreases binding to brain phosphatidylserine
K16I
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mutation moderately decreases binding to brain phosphatidylserine
Q10A
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mutation does not significantly decrease binding to brain phosphatidylserine
Q20A
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mutation moderately decreases binding to brain phosphatidylserine
R18P
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mutation strongly decreases binding to brain phosphatidylserine
R68A
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mutation strongly decreases binding to brain phosphatidylserine
S38A
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mutation does not significantly decrease binding to brain phosphatidylserine
V15S
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mutation strongly decreases binding to brain phosphatidylserine
Y14A
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mutation strongly decreases binding to brain phosphatidylserine
Y78A
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mutation does not significantly decrease binding to brain phosphatidylserine
I383S/Q385A
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mutation allow modification of Ras in the presence of UDP-N-acetyl-glucosamine and reduces the acceptance of UDP-glucose as a donor for glycosylation
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additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
medicine