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Literature summary extracted from
- Role of cysteine residues in pseudouridine synhases of different families (1999), Biochemistry, 38, 13106-13111.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.4.99.25 | - |
Escherichia coli |
| 5.4.99.28 | - |
Escherichia coli |
| 5.4.99.29 | - |
Escherichia coli |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.4.99.25 | C174A | kcat is 1.3fold higher than wild-type value, KM is 1.1fold lower than wild-type value | Escherichia coli |
| 5.4.99.25 | C193V | kcat is 1.5fold higher than wild-type value, KM is 1.5fold higher than wild-type value | Escherichia coli |
| 5.4.99.25 | C58A | kcat is 2.2fold higher than wild-type value, KM is 1.2fold higher than wild-type value | Escherichia coli |
| 5.4.99.25 | C58A/C174A/C193A | kcat is 1.1fold lower than wild-type value, KM is 1.2fold lower than wild-type value | Escherichia coli |
| 5.4.99.25 | additional information | substitution of cysteine for amino acids with nonnucleophilic side chains does not significantly alter the catalytic activity | Escherichia coli |
| 5.4.99.28 | C117A | kcat is 1.4fold lower than wild-type value, KM is 3.9fold higher than wild-type value | Escherichia coli |
| 5.4.99.28 | C117A/A128A | kcat is 1.3fold lower than wild-type value, KM is 1.5fold higher than wild-type value | Escherichia coli |
| 5.4.99.28 | C128A | kcat is 1.5fold higher than wild-type value, KM is 1.5fold higher than wild-type value | Escherichia coli |
| 5.4.99.28 | additional information | substitution of cysteine for amino acids with nonnucleophilic side chains does not significantly alter the catalytic activity | Escherichia coli |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.25 | 0.000124 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A/C174A/C193A | Escherichia coli | |
| 5.4.99.25 | 0.000127 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C174A | Escherichia coli | |
| 5.4.99.25 | 0.000146 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.25 | 0.000177 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A | Escherichia coli | |
| 5.4.99.25 | 0.000218 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C193V | Escherichia coli | |
| 5.4.99.28 | 0.000108 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.28 | 0.000157 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A | Escherichia coli | |
| 5.4.99.28 | 0.00016 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A | Escherichia coli | |
| 5.4.99.28 | 0.00042 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A | Escherichia coli |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.4.99.25 | Escherichia coli | P60340 | - |
- |
| 5.4.99.28 | Escherichia coli | P0AA37 | - |
- |
| 5.4.99.29 | Escherichia coli | P0AA37 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.4.99.25 | - |
Escherichia coli |
| 5.4.99.28 | - |
Escherichia coli |
| 5.4.99.29 | - |
Escherichia coli |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 5.4.99.25 | tRNA uridine55 = tRNA pseudouridine55 | RluA does not require thiol(ate) groups to effect catalysis | Escherichia coli | |
| 5.4.99.28 | tRNA uridine32 = tRNA pseudouridine32 | RluA does not require thiol(ate) groups to effect catalysis | Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.4.99.25 | tRNA uridine55 | substrate: Escherichia coli tRNAPhe | Escherichia coli | tRNA pseudouridine55 | - |
? |  |
| 5.4.99.28 | tRNA uridine32 | substrate: Escherichia coli tRNAPhe | Escherichia coli | tRNA pseudouridine32 | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.4.99.25 | TruB | - |
Escherichia coli |
| 5.4.99.28 | RluA | - |
Escherichia coli |
| 5.4.99.29 | RluA | - |
Escherichia coli |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.4.99.25 | 37 | - |
assay at | Escherichia coli |
| 5.4.99.28 | 37 | - |
assay at | Escherichia coli |
| 5.4.99.29 | 37 | - |
assay at | Escherichia coli |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.25 | 0.11 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A/C174A/C193A | Escherichia coli | |
| 5.4.99.25 | 0.12 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.25 | 0.16 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C174A | Escherichia coli | |
| 5.4.99.25 | 0.18 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C193V | Escherichia coli | |
| 5.4.99.25 | 0.26 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A | Escherichia coli | |
| 5.4.99.28 | 0.07 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A | Escherichia coli | |
| 5.4.99.28 | 0.074 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A | Escherichia coli | |
| 5.4.99.28 | 0.099 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.28 | 0.15 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A | Escherichia coli |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.4.99.25 | 7.5 | - |
assay at | Escherichia coli |
| 5.4.99.28 | 7.5 | - |
assay at | Escherichia coli |
| 5.4.99.29 | 7.5 | - |
assay at | Escherichia coli |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.25 | 821.9 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.25 | 825 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C193V | Escherichia coli | |
| 5.4.99.25 | 887 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A/C174A/C193A | Escherichia coli | |
| 5.4.99.25 | 1260 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C174A | Escherichia coli | |
| 5.4.99.25 | 1469 | - |
tRNA uridine55 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C58A | Escherichia coli | |
| 5.4.99.28 | 166.7 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A | Escherichia coli | |
| 5.4.99.28 | 471.3 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C117A/C128A | Escherichia coli | |
| 5.4.99.28 | 916.7 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe. wild-type enzyme | Escherichia coli | |
| 5.4.99.28 | 937.5 | - |
tRNA uridine32 | pH 7.5, 37°C, substrate: Escherichia coli tRNAPhe, mutant enzyme C128A | Escherichia coli |
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