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Literature summary for 2.1.1.56 extracted from

  • Zheng, S.; Shuman, S.
    Structure-function analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase (2008), RNA, 14, 696-705.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
His6-tagged proteins are expressed in Escherichia coli BL21(DE3) cells and in Saccharomyces cerevisiae strain YBS40 Vaccinia virus

Protein Variants

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 [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

Organism UniProt Comment Textmining
Vaccinia virus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA agarose column chromatography Vaccinia virus

Substrates and Products (Substrate)

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

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

Synonyms Comment Organism
cap methyltransferase
-
Vaccinia virus
RNA (guanine-N7) methyltransferase
-
Vaccinia virus

Turnover Number [1/s]

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