BRENDA - Enzyme Database

Electrostatic guidance of catalysis by a conserved glutamic acid in Escherichia coli dTMP synthase and bacteriophage T4 dCMP hydroxymethylase

Hardy, L.W.; Graves, K.L.; Nalivaika, E.; Biochemistry 34, 8422-8432 (1995)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.2.8
overexpression in Escherichia coli BL21; structural gene 42 encodes enzyme
Escherichia virus T4
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.1.2.8
E60D
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
2.1.2.8
E60Q
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.2.8
5-deazatetrahydrofolate
1 mM, 54% inhibition
Escherichia virus T4
2.1.2.8
5-fluorodeoxyuridylate
mechanism-based inhibitor
Escherichia virus T4
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.2.8
28487
-
2 * 28487
Escherichia virus T4
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Escherichia virus T4
-
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
Escherichia virus T4
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.2.8
Escherichia virus T4
-
Escherichia coli BL21 infected with bacteriophage T4
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.2.8
purification of wild-type enzyme and mutants E60Q and E60D
Escherichia virus T4
Reaction
EC Number
Reaction
Commentary
Organism
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
catalytic mechanism; ordered bi-bi mechanism
Escherichia virus T4
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
-
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
485791
Escherichia virus T4
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
role of active site Glu-60 is to accelerate bond cleavage between N5 of tetrahydrofolate and the methylene being transferred
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485791
Escherichia virus T4
r
Subunits
EC Number
Subunits
Commentary
Organism
2.1.2.8
homodimer
2 * 28487
Escherichia virus T4
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.2.8
0.0008
-
deoxycytidylate
mutant E60Q
Escherichia virus T4
2.1.2.8
0.007
-
deoxycytidylate
mutant E60D
Escherichia virus T4
2.1.2.8
12.7
-
deoxycytidylate
wild-type enzyme
Escherichia virus T4
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.1.2.8
5,10-methylenetetrahydrofolate
-
Escherichia virus T4
2.1.2.8
tetrahydrofolate
-
Escherichia virus T4
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.1.2.8
0.11
-
5-deazatetrahydrofolate
-
Escherichia virus T4
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.2.8
overexpression in Escherichia coli BL21; structural gene 42 encodes enzyme
Escherichia virus T4
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.2.8
5,10-methylenetetrahydrofolate
-
Escherichia virus T4
2.1.2.8
tetrahydrofolate
-
Escherichia virus T4
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.1.2.8
E60D
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
2.1.2.8
E60Q
mutant with decreased enzyme activity, impaired enzyme function in vitro and in vivo, uncoupling of early steps from later steps in catalysis
Escherichia virus T4
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.2.8
5-deazatetrahydrofolate
1 mM, 54% inhibition
Escherichia virus T4
2.1.2.8
5-fluorodeoxyuridylate
mechanism-based inhibitor
Escherichia virus T4
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.1.2.8
0.11
-
5-deazatetrahydrofolate
-
Escherichia virus T4
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.2.8
28487
-
2 * 28487
Escherichia virus T4
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Escherichia virus T4
-
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
Escherichia virus T4
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.2.8
purification of wild-type enzyme and mutants E60Q and E60D
Escherichia virus T4
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
-
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
involved in biosynthesis of DNA precursor
485791
Escherichia virus T4
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
role of active site Glu-60 is to accelerate bond cleavage between N5 of tetrahydrofolate and the methylene being transferred
485791
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485791
Escherichia virus T4
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.1.2.8
homodimer
2 * 28487
Escherichia virus T4
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.2.8
0.0008
-
deoxycytidylate
mutant E60Q
Escherichia virus T4
2.1.2.8
0.007
-
deoxycytidylate
mutant E60D
Escherichia virus T4
2.1.2.8
12.7
-
deoxycytidylate
wild-type enzyme
Escherichia virus T4