Literature summary for 2.1.2.8 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.

Search for more literature for 2.1.2.8

Cloned(Commentary)

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

Protein Variants

Protein Variants	Comment	Organism
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
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

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

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

Reaction

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

Substrates and Products (Substrate)

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

Subunits	Comment	Organism
homodimer	2 * 28487	Tequatrovirus T4

Turnover Number [1/s]

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

Cofactor

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

Ki Value [mM]

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

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Release 2023.1 (January 2023)