Literature summary for 2.1.2.8 extracted from

Lee, M.H.; Gautam-Basak, M.; Wooley, C.; Sander, E.G.
Deoxycytidylate hydroxymethylase: purification, properties, and the role of a thiol group in catalysis (1988), Biochemistry, 27, 1367-1373.

Search for more literature for 2.1.2.8

Activating Compound

Activating Compound	Comment	Organism	Structure
dithiothreitol	low specific activity samples can be activated by a 30 min room temperature incubation in 50 mM DTT	Tequatrovirus T4

Inhibitors

Inhibitors	Comment	Organism	Structure
5,5'-dithiobis(2-nitrobenzoic acid)	complete loss of activity, second-order kinetics, inactivation is reversed by dithiothreitol, inactivation involves the modification of one thiol group per mol of dimeric enzyme	Tequatrovirus T4
aminopterin	competitive inhibitor with respect to tetrahydrofolate	Tequatrovirus T4
dihydrofolate	competitive inhibitor with respect to tetrahydrofolate	Tequatrovirus T4
methotrexate	competitive inhibitor with respect to tetrahydrofolate	Tequatrovirus T4
N-ethylmaleimide	complete loss of activity, second-order kinetics, pH-dependent inactivation, dCMP protects against inactivation, dCMP plus either methotrexate or aminopterin greatly enhances protection, inactivation involves the modification of one thiol group per mol of dimeric enzyme	Tequatrovirus T4

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.04	-	tetrahydrofolate	-	Tequatrovirus T4

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
28000	-	2 * 28000, two equally sized subunits are arranged asymmetrically, SDS-PAGE	Tequatrovirus T4
51400	-	gel filtration	Tequatrovirus T4

Organism

Organism	UniProt	Comment	Textmining
Tequatrovirus T4	-	Escherichia coli strains Cr 63 and B infected with bacteriophage T4	-

Purification (Commentary)

Purification (Comment)	Organism
1026fold purification	Tequatrovirus T4
Escherichia coli B infected with	Tequatrovirus T4

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	-	Tequatrovirus T4

Storage Stability

Storage Stability	Organism
frozen, 20 mM KH2PO4, pH 6.8, 6 months	Tequatrovirus T4
room temperature, 24 h, stable	Tequatrovirus T4

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5,10-methylenetetrahydrofolate + H2O + deoxycytidylate	stoichiometry of dCMP binding to the dimeric enzyme is 1:1 and depends upon a reduced thiol group, one thiol-containing active site per mol of dimeric enzyme, sequential order of substrate binding	Tequatrovirus T4	tetrahydrofolate + 5-hydroxymethyldeoxycytidylate	-	r

Subunits

Subunits	Comment	Organism
dimer	2 * 28000, two equally sized subunits are arranged asymmetrically, SDS-PAGE	Tequatrovirus T4

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Tequatrovirus T4

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
20	-	room temperature, 24 h, stable	Tequatrovirus T4

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.6	-	assay at	Tequatrovirus T4

Cofactor

Cofactor	Comment	Organism	Structure
5,10-methylenetetrahydrofolate	active cofactor is likely 5,10-methylenetetrahydrofolate	Tequatrovirus T4
tetrahydrofolate	-	Tequatrovirus T4

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.001	-	aminopterin	-	Tequatrovirus T4
0.016	-	methotrexate	-	Tequatrovirus T4
0.021	-	dihydrofolate	-	Tequatrovirus T4

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Release 2023.1 (January 2023)