BRENDA - Enzyme Database

Evidence from 18O exchange studies for an exocyclic methylene intermediate in the reaction catalyzed by T4 deoxycytidylate hydroxymethylase

Butler, M.M.; Graves, K.L.; Hardy, L.W.; Biochemistry 33, 10521-10526 (1994)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.2.8
structural gene 42 encodes enzyme
Escherichia virus T4
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.1.2.8
C148G
mutant is unable to catalyze a detectable 18O exchange in either the absence or presence of the folate cofactor
Escherichia virus T4
2.1.2.8
D179N
mutant with altered substrate preference, favoring dUMP rather than dCMP
Escherichia virus T4
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.2.8
10-methyl-5,8-dideazatetrahydrofolate
competitive inhibitor with respect to tetrahydrofolate
Escherichia virus T4
2.1.2.8
5-deazatetrahydrofolate
competitive inhibitor with respect to tetrahydrofolate
Escherichia virus T4
2.1.2.8
5-fluorodeoxyuridylate
-
Escherichia virus T4
2.1.2.8
5-hydroxymethyl-dCMP
competitor of dCMP, inhibits formation of 5-hydroxymethyl-dCMP
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
5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4
Escherichia virus T4
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.2.8
Escherichia virus T4
-
Escherichia coli infected with bacteriophage T4
-
Reaction
EC Number
Reaction
Commentary
Organism
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
catalytic mechanism
Escherichia virus T4
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
2.1.2.8
3.1
-
dCMP formation
Escherichia virus T4
2.1.2.8
4.6
-
5-hydroxymethyl-dCMP formation
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
-
485790
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4
485790
Escherichia virus T4
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Cys-148 is catalytically essential
485790
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485790
Escherichia virus T4
r
2.1.2.8
additional information
enzyme also catalyzes 18O exchange from solvent water into 5-hydroxymethyl-dCMP in the absence of exogenous tetrahydrofolate and in the presence of tetrahydrofolate analogues that lack N-5, mechanism
485790
Escherichia virus T4
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
2.1.2.8
homodimer
-
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
0.01
-
10-methyl-5,8-dideazatetrahydrofolate
-
Escherichia virus T4
2.1.2.8
0.1
-
5-hydroxymethyl-dCMP
-
Escherichia virus T4
2.1.2.8
0.11
-
5-deazatetrahydrofolate
-
Escherichia virus T4
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.2.8
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
C148G
mutant is unable to catalyze a detectable 18O exchange in either the absence or presence of the folate cofactor
Escherichia virus T4
2.1.2.8
D179N
mutant with altered substrate preference, favoring dUMP rather than dCMP
Escherichia virus T4
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.2.8
10-methyl-5,8-dideazatetrahydrofolate
competitive inhibitor with respect to tetrahydrofolate
Escherichia virus T4
2.1.2.8
5-deazatetrahydrofolate
competitive inhibitor with respect to tetrahydrofolate
Escherichia virus T4
2.1.2.8
5-fluorodeoxyuridylate
-
Escherichia virus T4
2.1.2.8
5-hydroxymethyl-dCMP
competitor of dCMP, inhibits formation of 5-hydroxymethyl-dCMP
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
0.01
-
10-methyl-5,8-dideazatetrahydrofolate
-
Escherichia virus T4
2.1.2.8
0.1
-
5-hydroxymethyl-dCMP
-
Escherichia virus T4
2.1.2.8
0.11
-
5-deazatetrahydrofolate
-
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
5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4
Escherichia virus T4
?
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
2.1.2.8
3.1
-
dCMP formation
Escherichia virus T4
2.1.2.8
4.6
-
5-hydroxymethyl-dCMP formation
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
-
485790
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4
485790
Escherichia virus T4
?
2.1.2.8
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
Cys-148 is catalytically essential
485790
Escherichia virus T4
tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
485790
Escherichia virus T4
r
2.1.2.8
additional information
enzyme also catalyzes 18O exchange from solvent water into 5-hydroxymethyl-dCMP in the absence of exogenous tetrahydrofolate and in the presence of tetrahydrofolate analogues that lack N-5, mechanism
485790
Escherichia virus T4
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.1.2.8
homodimer
-
Escherichia virus T4