Protein Variants | Comment | Organism |
---|---|---|
S641A | mutation reduces activity in presence of Mg2+ to 93% of the activity of the wild-type enzyme | Escherichia phage T7 |
Y639/S641A | mutation reduces activity in presence of Mg2+ to 89% of the activity of the wild-type enzyme | Escherichia phage T7 |
Y639C | mutation reduces activity in presence of Mg2+ to 7.5% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates versus deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 11 | Escherichia phage T7 |
Y639F | mutation reduces the catalytic specificity for ribonucleoside triphosphates vs deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme by a factor of 20 and largely eliminates the KM-difference between rNTPs and dNTPs. The remaining specificity factor of 4 is turnover-number-mediated and is nearly eliminated if Mn2+ is substituted for Mg2+ in the reaction. Mn2+ substitution does not significantly affect the Km difference between rNTPs and dNTPs | Escherichia phage T7 |
Y639H | mutation reduces activity in presence of Mg2+ to 3.7% of the activity of the wild-type enzyme | Escherichia phage T7 |
Y639L | mutation reduces activity in presence of Mg2+ to 43% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates versus deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 11 | Escherichia phage T7 |
Y639M | mutation reduces activity in presence of Mg2+ to 50% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates versus deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 5.5 | Escherichia phage T7 |
Y639Q | mutation reduces activity in presence of Mg2+ to 1% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates vs deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 4.5 | Escherichia phage T7 |
Y639T | mutation reduces activity in presence of Mg2+ to 1.3% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates versus deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 6.5 | Escherichia phage T7 |
Y639V | mutation reduces activity in presence of Mg2+ to 4.3% of the activity of the wild-type enzyme. The mutation reduces the catalytic specificity for ribonucleoside triphosphates versus deoxynucleoside triphosphates during transcript elongation, which is about 80 for the wild-type enzyme. The remaining specificity factor is 19 | Escherichia phage T7 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0103 | - |
rGTP | pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mn2+, wild-type enzyme | Escherichia phage T7 | |
0.0104 | - |
rGTP | pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F | Escherichia phage T7 | |
0.015 | - |
dGTP | pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F | Escherichia phage T7 | |
0.0153 | - |
rGTP | pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mn2+, mutant enzyme Y639F | Escherichia phage T7 | |
0.0175 | - |
rGTP | pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, wild-type enzyme | Escherichia phage T7 | |
0.025 | - |
dGTP | pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F | Escherichia phage T7 | |
0.036 | - |
rUTP | pH 8.0, 37°C, activation by Mn2+, wild-type enzyme | Escherichia phage T7 | |
0.041 | - |
rUTP | pH 8.0, 37°C, activation by Mg2+, wild-type enzyme | Escherichia phage T7 | |
0.21 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme S641A | Escherichia phage T7 | |
0.22 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F/S641A | Escherichia phage T7 | |
0.25 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme | Escherichia phage T7 | |
0.32 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F | Escherichia phage T7 | |
0.387 | - |
dGTP | pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation with Mn2+ | Escherichia phage T7 | |
0.75 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F | Escherichia phage T7 | |
0.85 | - |
dGTP | pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, wild-type enzyme | Escherichia phage T7 | |
0.88 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme | Escherichia phage T7 | |
1.1 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F/S641A | Escherichia phage T7 | |
1.2 | - |
dUTP | pH 8.0, 37°C, activation by Mg2+, wild-type enzyme | Escherichia phage T7 | |
1.4 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, wild-type enzyme | Escherichia phage T7 | |
1.7 | - |
dUTP | pH 8.0, 37°C, activation by Mg2+, wild-type enzymes | Escherichia phage T7 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia phage T7 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dGTP + RNAn | - |
Escherichia phage T7 | diphosphate + RNAn+1 | - |
? | |
dUTP + RNAn | - |
Escherichia phage T7 | diphosphate + RNAn+1 | - |
? | |
nucleoside triphosphate + RNAn | the catalytic specificity for ribonucleoside triphosphates vs. deoxynucleoside triphosphates during transcript elongation is 80 | Escherichia phage T7 | diphosphate + RNAn+1 | - |
? | |
nucleoside triphosphate + RNAn | the enzyme requires DNA as template | Escherichia phage T7 | diphosphate + RNAn+1 | - |
? | |
rGTP + RNAn | - |
Escherichia phage T7 | diphosphate + RNAn+1 | - |
? | |
rUTP + RNAn | - |
Escherichia phage T7 | diphosphate + RNAn+1 | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia phage T7 | |
0.1 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F/S641A | Escherichia phage T7 | |
0.22 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, wild-type enzyme | Escherichia phage T7 | |
0.25 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F/S641A | Escherichia phage T7 | |
0.26 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme | Escherichia phage T7 | |
0.28 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F | Escherichia phage T7 | |
0.32 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme S641A | Escherichia phage T7 | |
0.34 | - |
dGTP | pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F | Escherichia phage T7 | |
0.38 | - |
rGTP | pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme S641A | Escherichia phage T7 |