Any feedback?
Literature summary extracted from
- The glutamate switch of bacteriophage T7 DNA helicase: role in coupling nucleotide triphosphate (NTP) and DNA binding to NTP hydrolysis (2011), J. Biol. Chem., 286, 23113-23120.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.6.1.39 | single-stranded DNA | the presence of single-stranded DNA stimulates the rate of dTTP hydrolysis about 100fold in the case of wild type enzyme | Escherichia phage T7 |  |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.6.1.39 | expressed in Escherichia coli HMS-174(DE3) cells | Escherichia phage T7 |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.6.1.39 | E343D | the mutant does not hydrolyze dTTP | Escherichia phage T7 |
| 3.6.1.39 | E343N | the mutant does not hydrolyze dTTP | Escherichia phage T7 |
| 3.6.1.39 | E343Q | the mutant does not hydrolyze dTTP | Escherichia phage T7 |
| 3.6.1.39 | N468A | the mutant hydrolyzes dTTP at rates comparable with wild type enzyme. N468A hydrolyzes nearly 4fold less efficiently than the wild type enzyme in the absence of single-stranded DNA and is not stimulated by the presence of single-stranded DNA | Escherichia phage T7 |
| 3.6.1.39 | R493A/N468A | the mutant hydrolyzes dTTP at rates comparable with wild type enzyme and is not stimulated by the presence of single-stranded DNA | Escherichia phage T7 |
| 3.6.1.39 | R493Q | the mutant hydrolyzes dTTP at rates comparable with wild type enzyme and is not stimulated by the presence of single-stranded DNA | Escherichia phage T7 |
| 3.6.1.39 | R493Q | the mutant maintains only the basal level of dTTPase activity but does not exhibit activation in the dTTP hydrolysis activity in the presence of single-stranded DNA | Escherichia phage T7 |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.6.1.39 | 0.06 | - |
dTTP | mutant enzyme R493Q, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.07 | - |
dTTP | mutant enzyme R493A/N468A, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.07 | - |
dTTP | wild type enzyme, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 2.6 | - |
dTTP | wild type enzyme, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.6.1.39 | Escherichia phage T7 | - |
- |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.6.1.39 | dTTP + H2O | Gp4 binds, as a hexamer, to single-stranded DNA in the presence of dTTP and translocates in a 5' to 3' direction along the DNA strand using the energy derived from the hydrolysis of dTTP | Escherichia phage T7 | dTDP + phosphate | - |
? | |
| 3.6.1.39 | additional information | wild type gp4 helicase unwinds the duplex DNA | Escherichia phage T7 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.6.1.39 | hexamer or heptamer | in the presence of dTTP Gp4 oligomerizes to form hexamers and heptamers | Escherichia phage T7 |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.6.1.39 | dTTPase | - |
Escherichia phage T7 |
| 3.6.1.39 | gene 4 protein | - |
Escherichia phage T7 |
| 3.6.1.39 | Gp4 | - |
Escherichia phage T7 |
| 3.6.1.39 | T7 DNA helicase | - |
Escherichia phage T7 |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.6.1.39 | 0.05 | - |
dTTP | mutant enzyme N468A, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.05 | - |
dTTP | mutant enzyme R493A/N468A, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.06 | - |
dTTP | mutant enzyme N468A, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.08 | - |
dTTP | mutant enzyme R493Q, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.22 | - |
dTTP | mutant enzyme R493Q, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.22 | - |
dTTP | wild type enzyme, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 0.35 | - |
dTTP | mutant enzyme R493A/N468A, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 20 | - |
dTTP | wild type enzyme, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.6.1.39 | 3.8 | - |
dTTP | mutant enzyme R493Q, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 5 | - |
dTTP | mutant enzyme R493A/N468A, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 5.7 | - |
dTTP | wild type enzyme, in the absence of DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
| 3.6.1.39 | 7.7 | - |
dTTP | wild type enzyme, in the presence of M13 single-stranded DNA, in 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 10 mM dithiothreitol, 50 mM potassium glutamate, at 37°C | Escherichia phage T7 | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
