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Literature summary for 5.4.99.25 extracted from

  • Spedaliere, C.J.; Hamilton, C.S.; Mueller, E.G.
    Functional importance of motif I of pseudouridine synthases: mutagenesis of aligned lysine and proline residues (2000), Biochemistry, 39, 9459-9465.
    View publication on PubMed
Search for more literature for 5.4.99.25

Protein Variants

Protein Variants Comment Organism
K19M kcat is 8fold lower than wild-type value. KM-value is 11fold higher than wild-type value Escherichia coli
K19R kcat is 10fold lower than wild-type value. KM-value is 6fold higher than wild-type value Escherichia coli
additional information mutant enzymes with substitution of the nearly invariant lysine and proline residues in motif I of RluA display only very mild kinetic impairment. Substitution of the aligned lysine and proline residues reduces structural stability Escherichia coli
P20G kcat is 4.8fold lower than wild-type value. KM-value is 2.2fold higher than wild-type value Escherichia coli
P20L kcat is 2.1fold lower than wild-type value. KM-value is 3.5fold higher than wild-type value Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000148
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme Escherichia coli
0.000324
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20G Escherichia coli
0.000517
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20L Escherichia coli
0.0009
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19R Escherichia coli
0.00161
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19M Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P60340
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tRNA uridine55 substrate: Escherichia coli tRNAPhe Escherichia coli tRNA pseudouridine552
-
 ?

Synonyms

Synonyms Comment Organism
TruB
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
36
-
 Tm-value for mutant enzyme P20L is 35.9°C Escherichia coli
38
-
 Tm-value for mutant enzyme P20G is 38.3°C Escherichia coli
40
-
 Tm-value for wild-type enzyme is 40.2°C Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.011
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19R Escherichia coli
0.012
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19M Escherichia coli
0.11
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme Escherichia coli
0.23
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20G Escherichia coli
0.52
-
 tRNA uridine55 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20L Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7.43
-
 tRNA uridine32 pH 7.5, 37°C, Escherichia coli tRNAPhe, wild-type enzyme Escherichia coli
7.45
-
 tRNA uridine32 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19M Escherichia coli
12.2
-
 tRNA uridine32 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme K19R Escherichia coli
709.8
-
 tRNA uridine32 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20G Escherichia coli
1006
-
 tRNA uridine32 pH 7.5, 37°C, Escherichia coli tRNAPhe, mutant enzyme P20L Escherichia coli