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Literature summary for 2.4.1.B62 extracted from

  • 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 (2005), J. Biol. Chem., 280, 37833-37838.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
I383S mutation in toxin B, 67% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose Clostridioides difficile
I383S/Q385A 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 Paeniclostridium sordellii
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 Clostridioides difficile
Q385A mutation in toxin B, 58% of wild-type catalytic efficiency with substrate UDP-alpha-D-glucose Clostridioides difficile
S385/A387Q mutation reverses the donor specificity of alpha-toxin from UDP-N-acetylglucosamine to UDP-glucose Clostridium novyi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.004
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Clostridium novyi
0.004
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Paeniclostridium sordellii
0.006
-
UDP-alpha-D-glucose wild-type, pH 7.5, 37°C Clostridioides difficile
0.008
-
UDP-alpha-D-glucose mutant I383S, pH 7.5, 37°C Clostridioides difficile
0.008
-
UDP-alpha-D-glucose mutant Q385A, pH 7.5, 37°C Clostridioides difficile
0.018
-
UDP-alpha-D-glucose mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile
0.026
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile
0.052
-
UDP-N-acetyl-alpha-D-glucosamine mutant Q385A, pH 7.5, 37°C Clostridioides difficile
0.061
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S, pH 7.5, 37°C Clostridioides difficile
0.96
-
UDP-N-acetyl-alpha-D-glucosamine wild-type, pH 7.5, 37°C Clostridioides difficile

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-alpha-D-glucose + H-Ras Paeniclostridium sordellii
-
UDP + D-glucosyl-H-Ras
-
?
UDP-alpha-D-glucose + H-Ras Paeniclostridium sordellii 6018
-
UDP + D-glucosyl-H-Ras
-
?
UDP-alpha-D-glucose + RhoA Clostridium novyi no substrate for wild-type. Mutation S385/A387Q reverses the donor specificity of alpha-toxin from UDP-N-acetylglucosamine to UDP-glucose UDP + D-glucosyl-RhoA
-
?
UDP-alpha-D-glucose + RhoA Clostridium novyi 19402 no substrate for wild-type. Mutation S385/A387Q reverses the donor specificity of alpha-toxin from UDP-N-acetylglucosamine to UDP-glucose UDP + D-glucosyl-RhoA
-
?

Organism

Organism UniProt Comment Textmining
Clostridioides difficile P18177 toxin B
-
Clostridioides difficile VPI 10463 P18177 toxin B
-
Clostridium novyi Q46149 alpha-toxin; type A
-
Clostridium novyi 19402 Q46149 alpha-toxin; type A
-
Paeniclostridium sordellii Q46342 cytotoxin L
-
Paeniclostridium sordellii 6018 Q46342 cytotoxin L
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-alpha-D-glucose + H-Ras
-
Paeniclostridium sordellii UDP + D-glucosyl-H-Ras
-
?
UDP-alpha-D-glucose + H-Ras
-
Paeniclostridium sordellii 6018 UDP + D-glucosyl-H-Ras
-
?
UDP-alpha-D-glucose + RhoA
-
Clostridioides difficile UDP + D-glucosyl-RhoA
-
?
UDP-alpha-D-glucose + RhoA no substrate for wild-type. Mutation S385/A387Q reverses the donor specificity of alpha-toxin from UDP-N-acetylglucosamine to UDP-glucose Clostridium novyi UDP + D-glucosyl-RhoA
-
?
UDP-alpha-D-glucose + RhoA no substrate for wild-type. Mutation S385/A387Q reverses the donor specificity of alpha-toxin from UDP-N-acetylglucosamine to UDP-glucose Clostridium novyi 19402 UDP + D-glucosyl-RhoA
-
?
UDP-alpha-D-glucose + RhoA
-
Clostridioides difficile VPI 10463 UDP + D-glucosyl-RhoA
-
?
UDP-N-acetyl-alpha-D-glucosamine + H-Ras no substrate for wild-type, but substrate for mutant I383S/Q385A Paeniclostridium sordellii UDP + N-acetyl-alpha-D-glucosamine-H-Ras
-
?
UDP-N-acetyl-alpha-D-glucosamine + H-Ras no substrate for wild-type, but substrate for mutant I383S/Q385A Paeniclostridium sordellii 6018 UDP + N-acetyl-alpha-D-glucosamine-H-Ras
-
?
UDP-N-acetyl-alpha-D-glucosamine + RhoA wild-type shows negligible activity, mutant I383S/Q385A has catalytic activity on UDP-N-acetyl-D-glucosamine Clostridioides difficile UDP + N-acetyl-D-glucosamine-RhoA
-
?
UDP-N-acetyl-alpha-D-glucosamine + RhoA wild-type shows negligible activity, mutant I383S/Q385A has catalytic activity on UDP-N-acetyl-D-glucosamine Clostridioides difficile VPI 10463 UDP + N-acetyl-D-glucosamine-RhoA
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.12
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S, pH 7.5, 37°C Clostridioides difficile
0.14
-
UDP-N-acetyl-alpha-D-glucosamine mutant Q385A, pH 7.5, 37°C Clostridioides difficile
0.39
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Clostridium novyi
0.39
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Paeniclostridium sordellii
0.42
-
UDP-alpha-D-glucose mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile
0.45
-
UDP-alpha-D-glucose mutant Q385A, pH 7.5, 37°C Clostridioides difficile
0.5
-
UDP-alpha-D-glucose mutant I383S, pH 7.5, 37°C Clostridioides difficile
0.59
-
UDP-alpha-D-glucose wild-type, pH 7.5, 37°C Clostridioides difficile
0.66
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.9
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S, pH 7.5, 37°C Clostridioides difficile
2.8
-
UDP-N-acetyl-alpha-D-glucosamine mutant Q385A, pH 7.5, 37°C Clostridioides difficile
22.8
-
UDP-alpha-D-glucose mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile
25.3
-
UDP-N-acetyl-alpha-D-glucosamine mutant I383S/Q385A, pH 7.5, 37°C Clostridioides difficile
58.3
-
UDP-alpha-D-glucose mutant Q385A, pH 7.5, 37°C Clostridioides difficile
66.7
-
UDP-alpha-D-glucose mutant I383S, pH 7.5, 37°C Clostridioides difficile
91.7
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Clostridium novyi
91.7
-
UDP-alpha-D-glucose mutant S385/A387Q, pH 7.5, 37°C Paeniclostridium sordellii
97.2
-
UDP-alpha-D-glucose wild-type, pH 7.5, 37°C Clostridioides difficile