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Literature summary extracted from
- T4 endonuclease V, perspectives on catalysis (1994), Ann. N. Y. Acad. Sci., 726, 181-197.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.25.1 | expressed in Escherichia coli | Tequatrovirus T4 |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.25.1 | A26Q | not processive | Tequatrovirus T4 |
| 3.1.25.1 | A3K | substantial level of AP- and dimer specific nicking, not processive | Tequatrovirus T4 |
| 3.1.25.1 | A3L | substantial level of AP- and dimer specific nicking, not processive | Tequatrovirus T4 |
| 3.1.25.1 | A3Q | substantial level of AP- and dimer specific nicking, not processive | Tequatrovirus T4 |
| 3.1.25.1 | E23D | no detectable N-glycosylase activity on pyrimidine dimers, partial retention of AP-lyase activity | Tequatrovirus T4 |
| 3.1.25.1 | E23Q | no detectable N-glycosylase activity on pyrimidine dimers, loss of AP-lyase activity | Tequatrovirus T4 |
| 3.1.25.1 | G133D | impaired ability to bind dimer-containing DNA | Tequatrovirus T4 |
| 3.1.25.1 | K130G | low dimer-specific binding activity, AP-endonuclease function several times as efficient as that of the wild-type | Tequatrovirus T4 |
| 3.1.25.1 | K130H | low dimer-specific binding activity, wild-type lyase activity, reduced glycosylase activity | Tequatrovirus T4 |
| 3.1.25.1 | K33Q | in combination with A26Q, not processive | Tequatrovirus T4 |
| 3.1.25.1 | additional information | insertion of Gly between the first 2 amino acids, lack of dimer-specific glycosylase and nicking activity | Tequatrovirus T4 |
| 3.1.25.1 | T2P | lack of dimer-specific glycosylase and nicking activity | Tequatrovirus T4 |
| 3.1.25.1 | T2S | wild-type level of dimer-specific glycosylase and nicking activity | Tequatrovirus T4 |
| 3.1.25.1 | T2V | intermediate activity level of dimer-specific glycosylase and nicking activity | Tequatrovirus T4 |
| 3.1.25.1 | W128S | wild-type glycosylase but virtually no AP-lyase activity | Tequatrovirus T4 |
| 3.1.25.1 | Y129N | - |
Tequatrovirus T4 |
| 3.1.25.1 | Y131N | in combination with Y129N, wild-type lyase activity retained, diminished dimer-specific binding and N-glycosylase activity | Tequatrovirus T4 |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.1.25.1 | CH2O | total inhibition if 4.4 methylations per molecule are present | Tequatrovirus T4 |  |
| 3.1.25.1 | NaBH4 | leads to a covalently linked enzyme-substrate product | Tequatrovirus T4 | |
| 3.1.25.1 | NaCN | reacts with imino intermediate | Tequatrovirus T4 | |
| 3.1.25.1 | NaCNBH3 | total inhibition if 4.4 methylations per molecule are present | Tequatrovirus T4 | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.1.25.1 | 16000 | - |
- |
Tequatrovirus T4 |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.25.1 | DNA + H2O | Tequatrovirus T4 | - |
? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.25.1 | Tequatrovirus T4 | - |
- |
- |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 3.1.25.1 | side-chain modification | methylation, after treatment with CH2O and NaCNBH3, N-terminal alpha-amino group is preferentially methylated, glycosylase and endonuclease activity are reduced to 20% if 0.8 methylations are present per enzyme molecule, with 4.4 methyl groups per enzyme both nicking and DNA-binding are abolished | Tequatrovirus T4 |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.1.25.1 | - |
Tequatrovirus T4 |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | specific for cis-syn cyclobutane pyrimidine dimers | Tequatrovirus T4 | |
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | cleavage on the 5'-side of dimers | Tequatrovirus T4 | |
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | salt concentrations of more than 40 mM cause the endonuclease V to shift into a distributive search mechanism instead of the processive mechanism | Tequatrovirus T4 | |
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | 2-step mechanism, pyrimidine dimer-DNA glycosylase and an apurinic/apyrimidinic endonuclease | Tequatrovirus T4 | |
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | processive | Tequatrovirus T4 | |
| 3.1.25.1 | endonucleolytic cleavage near pyrimidine dimers to products with 5'-phosphate | beta-elimination mechanism, imino intermediate | Tequatrovirus T4 |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.25.1 | DNA + H2O | - |
Tequatrovirus T4 | ? | - |
? | |
| 3.1.25.1 | additional information | not: unirradiated or heat-denatured DNA | Tequatrovirus T4 | ? | - |
? | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.25.1 | 6 | 8.5 | pH-optimum of glycosylase activity | Tequatrovirus T4 |
| 3.1.25.1 | 6.5 | - |
pH-optimum of endonuclease activity | Tequatrovirus T4 |
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