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Literature summary extracted from

  • Hardy, L.W.; Graves, K.L.; Nalivaika, E.
    Electrostatic guidance of catalysis by a conserved glutamic acid in Escherichia coli dTMP synthase and bacteriophage T4 dCMP hydroxymethylase (1995), Biochemistry, 34, 8422-8432.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.2.8 overexpression in Escherichia coli BL21 Tequatrovirus T4
2.1.2.8 structural gene 42 encodes enzyme Tequatrovirus T4

Protein Variants

EC Number Protein Variants Comment 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 Tequatrovirus 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 Tequatrovirus T4

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.2.8 5-deazatetrahydrofolate 1 mM, 54% inhibition Tequatrovirus T4
2.1.2.8 5-fluorodeoxyuridylate mechanism-based inhibitor Tequatrovirus T4

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.2.8 28487
-
 2 * 28487 Tequatrovirus T4

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.2.8 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate Tequatrovirus T4
-
 tetrahydrofolate + 5-hydroxymethyldeoxycytidylate involved in biosynthesis of DNA precursor ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.2.8 Tequatrovirus T4
-
 Escherichia coli BL21 infected with bacteriophage T4
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.2.8 purification of wild-type enzyme and mutants E60Q and E60D Tequatrovirus T4

Reaction

EC Number Reaction Comment Organism Reaction ID
2.1.2.8 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate ordered bi-bi mechanism Tequatrovirus T4
a
2.1.2.8 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate catalytic mechanism Tequatrovirus T4
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.2.8 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate
-
 Tequatrovirus T4 tetrahydrofolate + 5-hydroxymethyldeoxycytidylate involved in biosynthesis of DNA precursor ?
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 Tequatrovirus T4 tetrahydrofolate + 5-hydroxymethyldeoxycytidylate
-
 r

Subunits

EC Number Subunits Comment Organism
2.1.2.8 homodimer 2 * 28487 Tequatrovirus T4

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.2.8 0.0008
-
 deoxycytidylate mutant E60Q Tequatrovirus T4
2.1.2.8 0.007
-
 deoxycytidylate mutant E60D Tequatrovirus T4
2.1.2.8 12.7
-
 deoxycytidylate wild-type enzyme Tequatrovirus T4

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.2.8 5,10-methylenetetrahydrofolate
-
 Tequatrovirus T4
2.1.2.8 tetrahydrofolate
-
 Tequatrovirus T4

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.1.2.8 0.11
-
 5-deazatetrahydrofolate
-
 Tequatrovirus T4