Cloned (Comment) | Organism |
---|---|
- |
Escherichia coli |
pseudouridine synthase I is cloned behind a T7 promoter and expressed in Escherichia coli to about 20% of total soluble proteins | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D60A | mutants binds tRNA but is catalytically inactive and fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60A | catalytically inactive, fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60E | mutants binds tRNA but is catalytically inactive and fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60E | catalytically inactive, fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60K | mutants binds tRNA but is catalytically inactive and fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60K | catalytically inactive, fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60N | mutants binds tRNA but is catalytically inactive and fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60N | catalytically inactive, fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60S | mutants binds tRNA but is catalytically inactive and fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
D60S | catalytically inactive, fails to form covalent complexes with fluorouracil-substituted tRNA | Escherichia coli |
additional information | all Asp60 mutants bind tRNA but are catalytically inactive and fail to form covalent complexes with fluorouracil-substituted tRNA. It is concluded that the conserved Asp60 is essential for pseudouridine synthase activity | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5-fluorouracil tRNAPhe | time-dependent inactivation of pseudouridine synthase I and formation of a covalent complex with the enzyme that involves the 5-fluorouracil monophosphate at position 39 | Escherichia coli | |
fluorouracil-substituted tRNA | causes a time-dependent inactivation of pseudouridine synthase I and forms a covalent complex with the enzyme that involves the fluorouracil-substituted UMP at position 39. Upon incubation of 100 nM pseudouridine synthase with 0.001 mM fluorouracil-substituted tRNA at 15°C prior to addition of substrate, there is a time-dependent inactivation of the enzyme with a half-life of 35 min | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00094 | - |
tRNA uridine39 | pH 8.0, 22°C | Escherichia coli | |
0.0018 | - |
tRNA uridine39 | pH 8.0, 15°C | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31000 | - |
x * 31000, SDS-PAGE | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Escherichia coli | P07649 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzymes D60A, D60S, D60E, D60N, and D60K | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
tRNA uridine39 | tRNAPhe from yeast, which contains a single target for tRNA pseudouridine synthase I at U39 | Escherichia coli | tRNA pseudouridine39 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 31000, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
tRNA pseudouridine synthase I | - |
Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
1 h, about 50% loss of activity | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
tRNA uridine39 | pH 8.0, 15°C | Escherichia coli | |
0.73 | - |
tRNA uridine39 | pH 8.0, 22°C | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0001 | - |
fluorouracil-substituted tRNA | - |
Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
100 | - |
tRNA uridine39 | pH 8.0, 15°C | Escherichia coli | |
777 | - |
tRNA uridine39 | pH 8.0, 22°C | Escherichia coli |