Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 5.4.99.12 extracted from

  • Huang, L.; Pookanjanatavip, M.; Gu, X.; Santi, D.V.
    A conserved aspartate of tRNA pseudouridine synthase is essential for activity and a probable nucleophilic catalyst (1998), Biochemistry, 37, 344-351.
    View publication on PubMed

Cloned(Commentary)

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

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

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
31000
-
x * 31000, SDS-PAGE Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Escherichia coli P07649
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzymes D60A, D60S, D60E, D60N, and D60K Escherichia coli

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 31000, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
tRNA pseudouridine synthase I
-
Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
22
-
1 h, about 50% loss of activity Escherichia coli

Turnover Number [1/s]

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 [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0001
-
fluorouracil-substituted tRNA
-
Escherichia coli

kcat/KM [mM/s]

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