Information on EC 2.4.1.B62 - small GTPase glucosyltransferase and Organism(s) Clostridioides difficile and UniProt Accession P18177

for references in articles please use BRENDA:EC2.4.1.B62
preliminary BRENDA-supplied EC number
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EC Tree
     2 Transferases
         2.4 Glycosyltransferases
             2.4.1 Hexosyltransferases
                2.4.1.B62 small GTPase glucosyltransferase
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This record set is specific for:
Clostridioides difficile
UNIPROT: P18177 not found.
Word Map
The taxonomic range for the selected organisms is: Clostridioides difficile
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
+
a small GTPase
=
+
D-glucosyl-[a small GTPase]
Synonyms
Clostridium sordellii lethal toxin, cytotoxin B, cytotoxin L, glucosyltransferase TcdA, glucosyltransferase TcdB, haemorrhagic toxin, hemorrhagic toxin, letal toxin, lethal toxin, lethal-toxin, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cytotoxin B
300787
-
glucosyltransferase TcdA
300790
-
glucosyltransferase TcdB
300787
-
SYSTEMATIC NAME
IUBMB Comments
UDP-alpha-D-glucose:small GTPase glucosyltransferase
-
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 + a small GTPase
UDP + D-glucosyl-[a small GTPase]
show the reaction diagram
UDP-alpha-D-glucose + murine Rho GTPase
UDP + D-glucosyl-[murine Rho GTPase]
show the reaction diagram
-
-
-
?
UDP-alpha-D-glucose + RhoA
UDP + D-glucosyl-RhoA
show the reaction diagram
-
-
-
?
UDP-N-acetyl-alpha-D-glucosamine + RhoA
UDP + N-acetyl-D-glucosamine-RhoA
show the reaction diagram
wild-type shows negligible activity, mutant I383S/Q385A has catalytic activity on UDP-N-acetyl-D-glucosamine
-
-
?
UDP-alpha-D-glucose + a small GTPase
UDP + D-glucosyl-[a small GTPase]
show the reaction diagram
-
-
-
?
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
-
-
-
?
UDP-alpha-D-glucose + human Ras-GTPase
UDP + D-glucosyl-[human Ras-GTPase]
show the reaction diagram
activity of toxin A in human epithelial colorectal adenocarcinoma Caco-2 cells
-
-
?
UDP-alpha-D-glucose + Rac1
UDP + D-glucosyl-Rac1
show the reaction diagram
UDP-alpha-D-glucose + Rap2A
UDP + D-glucosyl-Rap2A
show the reaction diagram
modification occurs in vitro and in cells
-
-
?
UDP-alpha-D-glucose + RhoA
UDP + D-glucosyl-RhoA
show the reaction diagram
additional information
?
-
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
UDP-alpha-D-glucose + a small GTPase
UDP + D-glucosyl-[a small GTPase]
show the reaction diagram
UDP-alpha-D-glucose + murine Rho GTPase
UDP + D-glucosyl-[murine Rho GTPase]
show the reaction diagram
-
-
-
?
UDP-alpha-D-glucose + a small GTPase
UDP + D-glucosyl-[a small GTPase]
show the reaction diagram
-
-
-
?
UDP-alpha-D-glucose + human Ras-GTPase
UDP + D-glucosyl-[human Ras-GTPase]
show the reaction diagram
activity of toxin A in human epithelial colorectal adenocarcinoma Caco-2 cells
-
-
?
UDP-alpha-D-glucose + Rap2A
UDP + D-glucosyl-Rap2A
show the reaction diagram
modification occurs in vitro and in cells
-
-
?
UDP-alpha-D-glucose + RhoA
UDP + D-glucosyl-RhoA
show the reaction diagram
-
-
-
?
additional information
?
-
enzyme glucosylates and inactivates small GTPases of the Rho or Ras families, culminating in cytotoxicity
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
in addition to divalent cations, costimulation by K+ is required
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.006 - 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
0.42 - 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
1.9 - 25.3
UDP-N-acetyl-alpha-D-glucosamine
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
malfunction
the deletion mutant TcdBD1756-1780 shows similar glucosyltransferase and cysteine protease activity, cellular binding, and pore formation to wild-type TcdB, but it fails to induce the glucosylation of Rho GTPase Rac1 of host cells. Moreover, TcdBD1756-1780 is rapidly degraded in the endosome of target cells, and therefore its intact glucosyltransferase domain is unable to translocate efficiently into host cytosol. The decrease in the alpha helical structure in the composition of TcdBD1756-1780 may be due to the deletion of AA 1756-1780. Domain structures of wild-type and mutant enzymes, overview. The deletion of region 1756-1780 might lead to locking of the TcdB in endosomes, resulting in a failure to deliver GTD
metabolism
cytotoxin B possesses an inherent cysteine protease activity, which is responsible for auto-cleavage of glucosylating toxins
physiological function
metabolism
cytotoxin A possesses an inherent cysteine protease activity, which is responsible for auto-cleavage of glucosylating toxins
physiological function
additional information
the toxin requires InsP6-dependent autocleavage for activation
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
TCDB_CLODI
2366
0
269712
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
105000
-
and 220000, PAGE of toxin B
220000
-
and 105000, PAGE. The 220000 Da subunit of toxin B from strain 8864 migrates slightly faster than the subunit from strain 10463
307000
-
x * 307000, SDS-PAGE
308000
x * 308000, calculated
43000
-
x * 220000, x * 43000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
multimer
-
x * 220000, x * 43000, SDS-PAGE
additional information
the toxins are multi-domain proteins containing at least four functional domains. The N-terminus of the toxin harbors the glucosyltransferase domain (GTD) that inactivates host Rho GTPases by glucosylation and a cysteine protease domain (CPD) responsible for autoprocessing. The C-terminus, consisting of combined repetitive oligopeptides (CROP), is predicted to be a receptor binding domain (RBD)
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
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
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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C698A
mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
D587N
mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
H653A
mutant is able to glucosylate Rac protein but does not display cytotoxicity. Mutant does not show autocatalytical activity
I383S
mutation in toxin B, 67% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
I383S/Q385A
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
mutation in toxin B, 58% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His6-tagged deletion mutants from Bacillus megaterium by nickel affinity chromatography
purification of toxin B from strain 8864 that does not produce toxin A
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene tcdB, recombinant expression of C-terminally His6-tagged deletion mutants in a Bacillus megaterium expression system
expression in Bacillus megaterium
expression in Escherichia coli
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
construction of highly optimized plasmids encoding the receptor-binding domains from TcdA and TcdB in which any putative N-linked glycosylation sites are altered to test the potential of DNA vaccination against Clostridium difficile-associated disease. In mice and nonhuman primates, vaccination induces significant levels of both anti-receptor-binding domain antibodies (blood and stool) and receptor-binding domain-specific antibody-secreting cells. Sera from immunized mice and nonhuman primates can detect receptor-binding domain protein from transfected cells, as well as neutralize purified toxins in an in vitro cytotoxicity assay. Mice that are immunized with plasmids or given nonhuman-primate sera are protected from a lethal challenge with purified TcdA and/or TcdB. Immunized mice are significantly protected when challenged with Clostridium difficile spores from homologous (VPI 10463) and heterologous, epidemic (UK1) strains
analysis
medicine
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kim, H.; Kim, W.H.; Kim, M.; Jeong, S.H.; Lee, K.
Evaluation of a rapid membrane enzyme immunoassay for the simultaneous detection of glutamate dehydrogenase and toxin for the diagnosis of Clostridium difficile infection
Ann. Lab. Med.
34
235-239
2014
Clostridioides difficile
Manually annotated by BRENDA team
Faust, C.; Ye, B.; Song, K.
The enzymatic domain of Clostridium difficile toxin A is located within its N-terminal region
Biochem. Biophys. Res. Commun.
251
100-105
1998
Clostridioides difficile (P16154)
Manually annotated by BRENDA team
Castagliuolo, I.; Sardina, M.; Brun, P.; DeRos, C.; Mastrotto, C.; Lovato, L.; Palu, G.
Clostridium difficile toxin A carboxyl-terminus peptide lacking ADP-ribosyltransferase activity acts as a mucosal adjuvant
Infect. Immun.
72
2827-2836
2004
Clostridioides difficile, Clostridioides difficile VPI 10463
Manually annotated by BRENDA team
Bobo, L.D.; El Feghaly, R.E.; Chen, Y.S.; Dubberke, E.R.; Han, Z.; Baker, A.H.; Li, J.; Burnham, C.A.; Haslam, D.B.
MAPK-activated protein kinase 2 contributes to Clostridium difficile-associated inflammation
Infect. Immun.
81
713-722
2013
Clostridioides difficile
Manually annotated by BRENDA team
Just, I.; Wilm, M.; Selzer, J.; Rex, G.; von Eichel-Streiber, C.; Mann, M.; Aktories, K.
The enterotoxin from Clostridium difficile (ToxA) monoglucosylates the Rho proteins
J. Biol. Chem.
270
13932-13936
1995
Clostridioides difficile (P16154)
Manually annotated by BRENDA team
Jank, T.; Reinert, D.; Giesemann, T.; Schulz, G.; Aktories, K.
Change of the donor substrate specificity of Clostridium difficile toxin B by site-directed mutagenesis
J. Biol. Chem.
280
37833-37838
2005
Clostridioides difficile (P18177), Clostridioides difficile, Clostridium novyi (Q46149), Clostridium novyi, Paeniclostridium sordellii (Q46342), Paeniclostridium sordellii, Paeniclostridium sordellii 6018 (Q46342), Clostridium novyi 19402 (Q46149), Clostridioides difficile VPI 10463 (P18177)
Manually annotated by BRENDA team
Egerer, M.; Giesemann, T.; Jank, T.; Satchell, K.J.; Aktories, K.
Auto-catalytic cleavage of Clostridium difficile toxins A and B depends on cysteine protease activity
J. Biol. Chem.
282
25314-25321
2007
Clostridioides difficile (P16154), Clostridioides difficile (P18177)
Manually annotated by BRENDA team
Pruitt, R.; Chumbler, N.; Rutherford, S.; Farrow, M.; Friedman, D.; Spiller, B.; Lacy, D.
Structural determinants of Clostridium difficile toxin A glucosyltransferase activity
J. Biol. Chem.
287
8013-8020
2012
Clostridioides difficile (Q189K5), Clostridioides difficile 630 (Q189K5)
Manually annotated by BRENDA team
Shoshan, M.; Florin, I.; Thelestam, M.
Activation of cellular phospholipase A2 by Clostridium difficile toxin B
J. Cell. Biochem.
52
116-124
1993
Clostridioides difficile
Manually annotated by BRENDA team
Just, I.; Selzer, J.; Von Eichel-Streiber, C.; Aktories, K.
The low molecular mass GTP-binding protein Rho is affected by toxin A from Clostridium difficile
J. Clin. Invest.
95
1026-1031
1995
Clostridioides difficile
Manually annotated by BRENDA team
Sharp, S.E.; Ruden, L.O.; Pohl, J.C.; Hatcher, P.A.; Jayne, L.M.; Ivie, W.M.
Evaluation of the C.Diff Quik Chek Complete Assay, a new glutamate dehydrogenase and A/B toxin combination lateral flow assay for use in rapid, simple diagnosis of Clostridium difficile disease
J. Clin. Microbiol.
48
2082-2086
2010
Clostridioides difficile
Manually annotated by BRENDA team
Mitchell, T.; Ketley, J.; Burdon, D.; Candy, D.; Stephen, J.
The effects of Clostridium difficile toxins A and B on membrane integrity and protein synthesis in intestinal cells in vivo and in vitro and in McCoy cells in vitro
J. Med. Microbiol.
23
205-210
1987
Clostridioides difficile
Manually annotated by BRENDA team
Torres, J.
Purification and characterisation of toxin B from a strain of Clostridium difficile that does not produce toxin A
J. Med. Microbiol.
35
40-44
1991
Clostridioides difficile, Clostridioides difficile 8864
Manually annotated by BRENDA team
Blake, J.; Mitsikosta, F.; Metcalfe, M.
Immunological detection and cytotoxic properties of toxins from toxin A-positive, toxin B-positive Clostridium difficile variants
J. Med. Microbiol.
53
197-205
2004
Clostridioides difficile
Manually annotated by BRENDA team
Nam, S.T.; Seok, H.; Kim, D.H.; Nam, H.J.; Kang, J.K.; Eom, J.H.; Lee, M.B.; Kim, S.K.; Park, M.J.; Chang, J.S.; Ha, E.M.; Shong, K.E.; Hwang, J.S.; Kim, H.
Clostridium difficile toxin A inhibits erythropoietin receptor-mediated colonocyte focal adhesion through inactivation of Janus Kinase-2
J. Microbiol. Biotechnol.
22
1629-1635
2012
Clostridioides difficile, Clostridioides difficile VPI 10463
Manually annotated by BRENDA team
Seok, H.; Nam, H.J.; Nam, S.T.; Kang, J.K.; Kim, S.K.; Chang, J.S.; Ha, E.M.; Park, Y.J.; Kim, H.
Clostridium difficile toxin A inhibits the kinase activity of extracellular signal-related kinases 1 and 2 through direct binding
J. Microbiol. Biotechnol.
22
170-175
2012
Clostridioides difficile, Clostridioides difficile VPI 10463
Manually annotated by BRENDA team
Reinert, D.; Jank, T.; Aktories, K.; Schulz, G.
Structural basis for the function of Clostridium difficile toxin B
J. Mol. Biol.
351
973-981
2005
Clostridioides difficile (P18177), Clostridioides difficile VPI 10463 (P18177)
Manually annotated by BRENDA team
Ko, S.H.; Jeon, J.I.; Kim, H.; Kim, Y.J.; Youn, J.; Kim, J.M.
Mitogen-activated protein kinase/IkappaB kinase/NF-kappaB-dependent and AP-1-independent CX3CL1 expression in intestinal epithelial cells stimulated with Clostridium difficile toxin A
J. Mol. Med.
92
411-427
2014
Clostridioides difficile
Manually annotated by BRENDA team
Savidge, T,C,; Urvil, P.;, Oezguen, N.; Ali, K.; Choudhury, A.; Acharya, V.; Pinchuk, I.; Torres, A.G., English, R.D.; Wiktorowicz, J.E.; Loeffelholz, M.; Kumar, R.; Shi, L.; Nie, W.; Braun, W.; Herman, B., Hausladen, A.; Feng, H.; Stamler, J.S.; Pothoulakis, C.
Host S-nitrosylation inhibits clostridial small molecule-activated glucosylating toxins
Nat. Med.
17
1136-1141
2011
Clostridioides difficile
Manually annotated by BRENDA team
Shen, A.; Lupardus, P.J.; Gersch, M.M.; Puri, A.W.; Albrow, V.E.; Garcia, K.C.; Bogyo, M.
Defining an allosteric circuit in the cysteine protease domain of Clostridium difficile toxins
Nat. Struct. Mol. Biol.
18
364-371
2011
Clostridioides difficile (Q189K3), Clostridioides difficile 630 (Q189K3)
Manually annotated by BRENDA team
Just, I.; Selzer, J.; Wilm, M.; von Eichel-Streiber, C.; Mann, M.; Aktories, K.
Glucosylation of Rho proteins by Clostridium difficile toxin B
Nature
375
500-503
1995
Clostridioides difficile
Manually annotated by BRENDA team
Vesenka, G.D.; Majumdar, A.P.; Dubick, M.A.; Lyerly, D.M.; Wilkins, T.D.; Silva, J.; Geokas, M.C.
Stimulation of enzyme secretion from isolated pancreatic acini by Clostridium difficile toxin B
Toxicol. Lett.
34
261-269
1986
Clostridioides difficile
Manually annotated by BRENDA team
Roberts, A.; Shone, C.
Modification of surface histidine residues abolishes the cytotoxic activity of Clostridium difficile toxin A
Toxicon
39
325-333
2000
Clostridioides difficile, Clostridioides difficile VPI 10463
Manually annotated by BRENDA team
Baliban, S.M.; Michael, A.; Shammassian, B.; Mudakha, S.; Khan, A.S.; Cocklin, S.; Zentner, I.; Latimer, B.P.; Bouillaut, L.; Hunter, M.; Marx, P.; Sardesai, N.Y.; Welles, S.L.; Jacobson, J.M.; Weiner, D.B.; Kutzler, M.A.
An optimized, synthetic DNA vaccine encoding the toxin A and toxin B receptor binding domains of Clostridium difficile induces protective antibody responses in vivo
Infect. Immun.
82
4080-4091
2014
Clostridioides difficile (P16154), Clostridioides difficile (P18177)
Manually annotated by BRENDA team
Kasendra, M.; Barrile, R.; Leuzzi, R.; Soriani, M.
Clostridium difficile toxins facilitate bacterial colonization by modulating the fence and gate function of colonic epithelium
J. Infect. Dis.
209
1095-1104
2014
Clostridioides difficile (P16154)
Manually annotated by BRENDA team
Genth, H.; Schelle, I.; Just, I.
Metal ion activation of Clostridium sordellii lethal toxin and Clostridium difficile toxin B
Toxins
8
109
2016
Clostridioides difficile (P18177), Clostridioides difficile, Paeniclostridium sordellii (V5T923), Paeniclostridium sordellii
Manually annotated by BRENDA team
Genth, H.; Junemann, J.; Laemmerhirt, C.M.; Luecke, A.C.; Schelle, I.; Just, I.; Gerhard, R.; Pich, A.
Difference in mono-O-glucosylation of Ras subtype GTPases between toxin A and toxin B from Clostridioides difficile strain 10463 and lethal toxin from Clostridium sordellii strain 6018
Front. Microbiol.
9
3078
2018
Clostridioides difficile (P16154), Clostridioides difficile (P18177), Clostridioides difficile, Paeniclostridium sordellii (Q46342), Paeniclostridium sordellii, Paeniclostridium sordellii 6018 (Q46342), Clostridioides difficile 10463 (P16154), Clostridioides difficile 10463 (P18177)
Manually annotated by BRENDA team
Chen, S.; Wang, H.; Gu, H.; Sun, C.; Li, S.; Feng, H.; Wang, J.
Identification of an essential region for translocation of Clostridium difficile toxin B
Toxins
8
241
2016
Clostridioides difficile (P18177)
Manually annotated by BRENDA team
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