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 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 | 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 | 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 | 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 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 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 |