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Literature summary for 2.1.2.8 extracted from
- Evidence from 18O exchange studies for an exocyclic methylene intermediate in the reaction catalyzed by T4 deoxycytidylate hydroxymethylase (1994), Biochemistry, 33, 10521-10526.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| structural gene 42 encodes enzyme | Tequatrovirus T4 |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C148G | mutant is unable to catalyze a detectable 18O exchange in either the absence or presence of the folate cofactor | Tequatrovirus T4 |
| D179N | mutant with altered substrate preference, favoring dUMP rather than dCMP | Tequatrovirus T4 |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 10-methyl-5,8-dideazatetrahydrofolate | competitive inhibitor with respect to tetrahydrofolate | Tequatrovirus T4 |  |
| 5-deazatetrahydrofolate | competitive inhibitor with respect to tetrahydrofolate | Tequatrovirus T4 | |
| 5-fluorodeoxyuridylate | - |
Tequatrovirus T4 | |
| 5-hydroxymethyl-dCMP | competitor of dCMP, inhibits formation of 5-hydroxymethyl-dCMP | 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 | 5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4 | ? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Tequatrovirus T4 | - |
Escherichia coli infected with bacteriophage T4 | - |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate = tetrahydrofolate + 5-hydroxymethyldeoxycytidylate | catalytic mechanism | Tequatrovirus T4 |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 3.1 | - |
dCMP formation | Tequatrovirus T4 |
| 4.6 | - |
5-hydroxymethyl-dCMP formation | Tequatrovirus T4 |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate | - |
Tequatrovirus T4 | tetrahydrofolate + 5-hydroxymethyldeoxycytidylate | 5-hydroxymethyl-dCMP is essential for DNA synthesis in phage T4 | ? | |
| 5,10-methylenetetrahydrofolate + H2O + deoxycytidylate | Cys-148 is catalytically essential | Tequatrovirus T4 | tetrahydrofolate + 5-hydroxymethyldeoxycytidylate | - |
r | |
| 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 | Tequatrovirus T4 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | - |
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 |
|---|---|---|---|---|---|
| 0.01 | - |
10-methyl-5,8-dideazatetrahydrofolate | - |
Tequatrovirus T4 | |
| 0.1 | - |
5-hydroxymethyl-dCMP | - |
Tequatrovirus T4 | |
| 0.11 | - |
5-deazatetrahydrofolate | - |
Tequatrovirus T4 | |
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