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Literature summary extracted from

  • Huang, Y.; Beaudry, A.; McSwiggen, J.; Sousa, R.
    Determinants of ribose specificity in RNA polymerization: effects of Mn2+ and deoxynucleoside monophosphate incorporation into transcripts (1997), Biochemistry, 36, 13718-13728.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.6 S641A mutation reduces activity in presence of Mg2+ to 93% of the activity of the wild-type enzyme Escherichia phage T7
2.7.7.6 Y639/S641A mutation reduces activity in presence of Mg2+ to 89% of the activity of the wild-type enzyme Escherichia phage T7
2.7.7.6 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
2.7.7.6 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
2.7.7.6 Y639H mutation reduces activity in presence of Mg2+ to 3.7% of the activity of the wild-type enzyme Escherichia phage T7
2.7.7.6 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
2.7.7.6 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
2.7.7.6 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
2.7.7.6 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
2.7.7.6 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.6 0.0103
-
 rGTP pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mn2+, wild-type enzyme Escherichia phage T7
2.7.7.6 0.0104
-
 rGTP pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.015
-
 dGTP pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.0153
-
 rGTP pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mn2+, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.0175
-
 rGTP pH 8.0, 37°C, rGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, wild-type enzyme Escherichia phage T7
2.7.7.6 0.025
-
 dGTP pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.036
-
 rUTP pH 8.0, 37°C, activation by Mn2+, wild-type enzyme Escherichia phage T7
2.7.7.6 0.041
-
 rUTP pH 8.0, 37°C, activation by Mg2+, wild-type enzyme Escherichia phage T7
2.7.7.6 0.21
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme S641A Escherichia phage T7
2.7.7.6 0.22
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F/S641A Escherichia phage T7
2.7.7.6 0.25
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme Escherichia phage T7
2.7.7.6 0.32
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.387
-
 dGTP pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation with Mn2+ Escherichia phage T7
2.7.7.6 0.75
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.85
-
 dGTP pH 8.0, 37°C, dGTP as elongation substrate during dinucleotide synthesis, activation by Mg2+, wild-type enzyme Escherichia phage T7
2.7.7.6 0.88
-
 dGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme Escherichia phage T7
2.7.7.6 1.1
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F/S641A Escherichia phage T7
2.7.7.6 1.2
-
 dUTP pH 8.0, 37°C, activation by Mg2+, wild-type enzyme Escherichia phage T7
2.7.7.6 1.4
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, wild-type enzyme Escherichia phage T7
2.7.7.6 1.7
-
 dUTP pH 8.0, 37°C, activation by Mg2+, wild-type enzymes Escherichia phage T7

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.6 Escherichia phage T7
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.6 dGTP + RNAn
-
 Escherichia phage T7 diphosphate + RNAn+1
-
 ?
2.7.7.6 dUTP + RNAn
-
 Escherichia phage T7 diphosphate + RNAn+1
-
 ?
2.7.7.6 nucleoside triphosphate + RNAn the catalytic specificity for ribonucleoside triphosphates vs. deoxynucleoside triphosphates during transcript elongation is 80 Escherichia phage T7 diphosphate + RNAn+1
-
 ?
2.7.7.6 nucleoside triphosphate + RNAn the enzyme requires DNA as template Escherichia phage T7 diphosphate + RNAn+1
-
 ?
2.7.7.6 rGTP + RNAn
-
 Escherichia phage T7 diphosphate + RNAn+1
-
 ?
2.7.7.6 rUTP + RNAn
-
 Escherichia phage T7 diphosphate + RNAn+1
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.6 additional information
-
 additional information
-
 Escherichia phage T7
2.7.7.6 0.1
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F/S641A Escherichia phage T7
2.7.7.6 0.22
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, wild-type enzyme Escherichia phage T7
2.7.7.6 0.25
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F/S641A Escherichia phage T7
2.7.7.6 0.26
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, wild-type enzyme Escherichia phage T7
2.7.7.6 0.28
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.32
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme S641A Escherichia phage T7
2.7.7.6 0.34
-
 dGTP pH 8.0, 37°C, initiating nucleotide in dGrA synthesis, mutant enzyme Y639F Escherichia phage T7
2.7.7.6 0.38
-
 rGTP pH 8.0, 37°C, initiating nucleotide in rGrA synthesis, mutant enzyme S641A Escherichia phage T7