Cloned (Comment) | Organism |
---|---|
His6-tagged proteins are expressed in Escherichia coli BL21(DE3) cells and in Saccharomyces cerevisiae strain YBS40 | Vaccinia virus |
Protein Variants | Comment | Organism |
---|---|---|
D598E | the mutant enzyme has 1% of wild type activity | Vaccinia virus |
D598N | the mutant enzyme hasless than 1% of wild type activity | Vaccinia virus |
D604E | the mutant enzyme has 112% of wild type activity | Vaccinia virus |
D604N | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
D620E | the mutant enzyme has 59% of wild type activity | Vaccinia virus |
D620N | the mutant enzyme has 11% of wild type activity | Vaccinia virus |
D657A | the mutant enzyme has 112% of wild type activity | Vaccinia virus |
D676A | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
D676E | the mutant enzyme has 40% of wild type activity | Vaccinia virus |
D676N | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
E763A | the mutant enzyme has 16% of wild type activity | Vaccinia virus |
E763D | the mutant enzyme has 1% of wild type activity | Vaccinia virus |
E763Q | the mutant enzyme has 32% of wild type activity | Vaccinia virus |
E816A | the mutant enzyme has 76% of wild type activity | Vaccinia virus |
F609A | the mutant enzyme has 117% of wild type activity | Vaccinia virus |
F659A | the mutant enzyme has 96% of wild type activity | Vaccinia virus |
F679A | the mutant enzyme has 5% of wild type activity | Vaccinia virus |
F679H | the mutant enzyme has 13% of wild type activity | Vaccinia virus |
F679I | the mutant enzyme has 18% of wild type activity | Vaccinia virus |
F679L | the mutant enzyme has 55% of wild type activity | Vaccinia virus |
F679N | the mutant enzyme has 18% of wild type activity | Vaccinia virus |
F679V | the mutant enzyme has 20% of wild type activity | Vaccinia virus |
F814A | the mutant enzyme has 99% of wild type activity | Vaccinia virus |
F815A | the mutant enzyme has 92% of wild type activity | Vaccinia virus |
H682A | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
H682N | the mutant enzyme has 1% of wild type activity | Vaccinia virus |
H682Q | the mutant enzyme has 4% of wild type activity | Vaccinia virus |
K573A | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
K573Q | the mutant enzyme has less than 1% of wild type activity | Vaccinia virus |
K573R | the mutant enzyme has 1% of wild type activity | Vaccinia virus |
K607A | the mutant enzyme has101% of wild type activity | Vaccinia virus |
K607Q | the mutant enzyme has 74% of wild type activity | Vaccinia virus |
K607R | the mutant enzyme has 13% of wild type activity | Vaccinia virus |
N570A | the mutant enzyme has 37% of wild type activity | Vaccinia virus |
N570D | the mutant enzyme has 21% of wild type activity | Vaccinia virus |
N570Q | the mutant enzyme has 73% of wild type activity | Vaccinia virus |
N601A | the mutant enzyme has 32% of wild type activity | Vaccinia virus |
Q678A | the mutant enzyme has 55% of wild type activity | Vaccinia virus |
R632K | the mutant enzyme has 72% of wild type activity | Vaccinia virus |
R632Q | the mutant enzyme has 58% of wild type activity | Vaccinia virus |
S569A | the mutant enzyme has 39% of wild type activity | Vaccinia virus |
V750A | the mutant enzyme has 59% of wild type activity | Vaccinia virus |
W677A | the mutant enzyme has 4% of wild type activity | Vaccinia virus |
Y608A | the mutant enzyme has 3% of wild type activity | Vaccinia virus |
Y608F | the mutant enzyme has 74% of wild type activity | Vaccinia virus |
Y608L | the mutant enzyme has 62% of wild type activity | Vaccinia virus |
Y608S | the mutant enzyme has 2% of wild type activity | Vaccinia virus |
Y764A | the mutant enzyme has 18% of wild type activity | Vaccinia virus |
Y836A | the mutant enzyme has 15% of wild type activity | Vaccinia virus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0075 | - |
S-adenosyl-L-methionine | wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.008 | - |
S-adenosyl-L-methionine | mutant enzyme K607A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0095 | - |
S-adenosyl-L-methionine | mutant enzyme N570A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.023 | - |
G(5')pppR-RNA | wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.031 | - |
G(5')pppR-RNA | mutant enzyme K607A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.05 | - |
G(5')pppR-RNA | mutant enzyme F679L, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.074 | - |
G(5')pppR-RNA | mutant enzyme N570A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.143 | - |
G(5')pppR-RNA | mutant enzyme F679N, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.176 | - |
G(5')pppR-RNA | mutant enzyme F679I, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.183 | - |
G(5')pppR-RNA | mutant enzyme R632Q, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.2 | - |
G(5')pppR-RNA | mutant enzyme K607Q, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.218 | - |
G(5')pppR-RNA | mutant enzyme E763A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.255 | - |
G(5')pppR-RNA | mutant enzyme F679H, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.293 | - |
G(5')pppR-RNA | mutant enzyme F679V, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.314 | - |
G(5')pppR-RNA | mutant enzyme K607R, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.446 | - |
G(5')pppR-RNA | mutant enzyme N570D, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vaccinia virus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-NTA agarose column chromatography | Vaccinia virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + G(5')pppR-RNA | - |
Vaccinia virus | S-adenosyl-L-homocysteine + m7G(5')pppR-RNA | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | the guanine-N7 methyltransferase domain of vaccinia virus mRNA capping enzyme is a heterodimer composed of a catalytic subunit and a stimulatory subunit, X-ray crystallography | Vaccinia virus |
Synonyms | Comment | Organism |
---|---|---|
cap methyltransferase | - |
Vaccinia virus |
RNA (guanine-N7) methyltransferase | - |
Vaccinia virus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0018 | - |
G(5')pppR-RNA | mutant enzyme K607R, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0022 | - |
G(5')pppR-RNA | mutant enzyme E763A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.003 | - |
G(5')pppR-RNA | mutant enzyme F679H, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.003 | - |
G(5')pppR-RNA | mutant enzyme N570D, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0035 | - |
G(5')pppR-RNA | mutant enzyme F679N, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.004 | - |
G(5')pppR-RNA | mutant enzyme F679I, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.004 | - |
G(5')pppR-RNA | mutant enzyme F679V, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0057 | - |
G(5')pppR-RNA | mutant enzyme N570A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.007 | - |
S-adenosyl-L-methionine | mutant enzyme N570A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0073 | - |
G(5')pppR-RNA | mutant enzyme R632Q, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.0095 | - |
G(5')pppR-RNA | mutant enzyme F679L, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.016 | - |
S-adenosyl-L-methionine | mutant enzyme K607A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.018 | - |
G(5')pppR-RNA | mutant enzyme K607Q, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.02 | - |
G(5')pppR-RNA | mutant enzyme K607A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.022 | - |
G(5')pppR-RNA | mutant enzyme E763A, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.022 | - |
S-adenosyl-L-methionine | wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus | |
0.022 | - |
G(5')pppR-RNA | wild type enzyme, in 50 mM Tris-HCl (pH 7.5), 5 mM dithiothreitol, at 30°C | Vaccinia virus |