Information on EC 2.7.7.50 - mRNA guanylyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.7.7.50
-
RECOMMENDED NAME
GeneOntology No.
mRNA guanylyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GTP + (5')ppPur-mRNA = diphosphate + G(5')pppPur-mRNA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nucleotidyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
mRNA capping I
-
-
SYSTEMATIC NAME
IUBMB Comments
GTP:mRNA guanylyltransferase
The enzyme can also modify synthetic poly(A) and poly(G) to form the structures m7G(5')pppAn and m7G(5')pppGn.
CAS REGISTRY NUMBER
COMMENTARY hide
56941-23-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
a giant virus of amoeba, e.g. isolated parasite of Acanthamoeba polyphaga
Uniprot
Manually annotated by BRENDA team
i.e. baculovirus; LEF-4 subunit of DNA-dependent RNA-polymerase
-
-
Manually annotated by BRENDA team
l3 genome segment, protein lambdaC; strain 1733
-
-
Manually annotated by BRENDA team
strain 1733
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
bluetongue virus serotype 10
from infected Spodoptera frugiperda cells via baculovirus; i.e. BTV serotype 10; mino core protein VP4
-
-
Manually annotated by BRENDA team
calf
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
CMA035C
-
-
Manually annotated by BRENDA team
viral polyprotein
UniProt
Manually annotated by BRENDA team
viral polyprotein
UniProt
Manually annotated by BRENDA team
Orf3p gene product
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
mRNA-capping enzyme subunit alpha
UniProt
Manually annotated by BRENDA team
mRNA-capping enzyme subunit alpha
UniProt
Manually annotated by BRENDA team
mRNA-capping enzyme subunit alpha
UniProt
Manually annotated by BRENDA team
from infected Nicotiana tabacum leaves; i.e. TMV
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
mRNA-capping enzyme catalytic subunit
UniProt
Manually annotated by BRENDA team
mRNA-capping enzyme catalytic subunit
UniProt
Manually annotated by BRENDA team
strain WR
-
-
Manually annotated by BRENDA team
viral polyprotein
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-fluoro-2'-deoxy-GTP + pp(5')RNA
2'-fluoro-2'-deoxy-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.16fold efficiency compared with substrate GTP
-
-
?
2-amino-6-chloropurine-beta-D-ribose triphosphate + pp(5')RNA
2-amino-6-chloropurine-beta-D-ribose-(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.08fold efficiency compared with substrate GTP
-
-
?
3'-deoxy-GTP + pp(5')RNA
3'-deoxy-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.15fold efficiency compared with substrate GTP
-
-
?
3'-O-methyl-GTP + pp(5')RNA
3'-O-methyl-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.38fold efficiency compared with substrate GTP
-
-
?
6-methylthio-GTP + pp(5')RNA
6-methylthio-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.07fold efficiency compared with substrate GTP
-
-
?
6-thio-GTP + pp(5')RNA
6-thio-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.16fold efficiency compared with substrate GTP
-
-
?
8-bromo-GTP + pp(5')RNA
8-bromo-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.40fold efficiency compared with substrate GTP
-
-
?
8-iodoGTP + pp(5')RNA
8-iodo-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
1.0fold efficiency compared with substrate GTP
-
-
?
8-[(6-amino)hexyl]-amino-GTP-ATTO-680 + pp(5')RNA
G(5')ppp(5')RNA + diphosphate
show the reaction diagram
fluorescent substate analog
-
-
?
dGTP + pp(5')RNA
dG(5')ppp(5')RNA + diphosphate
show the reaction diagram
GTP + (5')ppPur-mRNA
diphosphate + G(5')pppPur-mRNA
show the reaction diagram
GTP + pp(5')ApGp
G(5')ppp(5')ApGp + diphosphate
show the reaction diagram
GTP + pp(5')GpCpC
G(5')ppp(5')GpCpC + diphosphate
show the reaction diagram
-
-
guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue
r
GTP + pp(5')GpXp
G(5')ppp(5')GpXp + diphosphate
show the reaction diagram
bluetongue virus serotype 10
-
-
guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue
r
GTP + pp(5')RNA
G(5')ppp(5')RNA + diphosphate
show the reaction diagram
GTP + ppApG
G(5')ppp(5')ApG + diphosphate
show the reaction diagram
-
-
guanosine residue linked 5' through three phosphates to the 5' position of the terminal residue
r
GTP + ppGpC
G(5')ppp(5')GpC + diphosphate
show the reaction diagram
GTP + ppp(5')ApG
G(5')pppp(5')ApG + diphosphate
show the reaction diagram
GTP + ppp(5')ApGp
G(5')pppp(5')ApGp + diphosphate
show the reaction diagram
-
-
guanosine residue linked 5' through four phosphates to the 5' position of the terminal residue
?
GTP + ppp(5')RNA
G(5')pppp(5')RNA + diphosphate
show the reaction diagram
GTP + pppGpCpC
G(5')pppp(5')GpCpC + diphosphate
show the reaction diagram
-
-
guanosine residue linked 5' through four phosphates to the 5' position of the terminal residue
r
GTPgammaS + pp(5')RNA
?
show the reaction diagram
ITP + pp(5')RNA
I(5')ppp(5')RNA + diphosphate
show the reaction diagram
N1-methyl-GTP + pp(5')RNA
N1-methyl-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
1.1fold efficiency compared with substrate GTP
-
-
?
NTP + H2O
NDP + phosphate
show the reaction diagram
-
-
-
-
?
O6-methyl-GTP + pp(5')RNA
O6-methyl-G(5')ppp(5')RNA + diphosphate
show the reaction diagram
-
0.48fold efficiency compared with substrate GTP
-
-
?
pppG + ppp(5')A(pA)n
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
GTP + (5')ppPur-mRNA
diphosphate + G(5')pppPur-mRNA
show the reaction diagram
GTP + pp(5')RNA
G(5')ppp(5')RNA + diphosphate
show the reaction diagram
GTP + ppp(5')RNA
G(5')pppp(5')RNA + diphosphate
show the reaction diagram
NTP + H2O
NDP + phosphate
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
can partially replace Mg2+ in activation
Co2+
30% of the activity with Mg2+
K+
-
stimulates, maximal activation at 6 mM KCl
Na+
-
stimulates, maximal activation at 50-75 mM NaCl
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2'-fluoro-2'-deoxy-GTP
-
-
2-amino-6-chloropurine-ribose triphosphate
-
-
-
3-[(5Z)-5-(2,3-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
3-[(5Z)-5-(2,4-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
3-[(5Z)-5-(4-tert-butylbenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
Co2+
-
above 0.1 mM
diphosphate
mizoribine 5'-phosphate
-
compound can inhibit the formation of the RNA cap structure catalyzed by human capping enzyme. In the presence of mizoribine 5'-phosphate, the RNA 5'-triphosphatase activity appears to be relatively unaffected while the RNA guanylyltransferase activity is inhibited. Mizoribine 5'-phosphate is a non-competitive inhibitor that likely targets an allosteric site
Mn2+
-
can partially replace Mg2+ in activation, inhibits in presence of Mg2+
N-ethylmaleimide
-
-
NaCl
-
90% inhibition at 0.3 M
phosphate
ribavirin triphosphate
-
concentration 200fold more than the GTP input, 50% inhibition
sodium phosphonoformate
-
foscarnet, inhibition of the formation of the covalent intermediate at 5 mM
tetrapotassium diphosphate
-
inhibition of the formation of the covalent intermediate at 5 mM
Zn2+
-
above 10 mM
[(5Z)-4-oxo-5-(4-phenoxybenzylidene)-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
[(5Z)-5-(3,5-diiodo-2-methoxybenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
[(5Z)-5-[4-(benzyloxy)benzylidene]-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bovine serum albumin
-
stimulates
-
RNA 5'-triphosphatase
-
binding in the capping enzyme complex stimulates the mRNA guanylyltransferase by 10fold, icreases affinity for GTP
-
S-adenosylmethionine
-
stimulates up to 2fold
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
diphosphate
-
pH 7.0, 37°C
0.019
diphosphate terminated poly(A) with an average chain length of 2000 nucleotides
-
-
-
0.000019
diphosphate-ended poly(A)
-
pH 7.5, 37°C
-
0.0011 - 0.017
GTP
0.00014
lambdac17RNA
-
pH 7.5, 37°C
-
0.000014
pp(5')A(pA)n
-
pH 7.9, 37°C
-
0.000285
pp(5')ApGp
-
0.0005 - 0.004
pp(5')GCC(A2,U2G)n
-
0.00025 - 0.0003
RNA
0.0002
termini of 5'-triphosphate poly(A)
-
pH 7.8, 37°C
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0233 - 0.0267
GTP
0.0147 - 0.0187
RNA
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0099 - 0.016
3-[(5Z)-5-(2,3-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.009 - 0.0098
3-[(5Z)-5-(2,4-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.0075 - 0.0095
3-[(5Z)-5-(4-tert-butylbenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.004 - 0.0077
[(5Z)-4-oxo-5-(4-phenoxybenzylidene)-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
0.0015 - 0.0046
[(5Z)-5-(3,5-diiodo-2-methoxybenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
0.0029 - 0.0044
[(5Z)-5-[4-(benzyloxy)benzylidene]-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
additional information
additional information
-
inhibition kinetic analysis of both steps of forward and reverse reactions, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.4
2'-fluoro-2'-deoxy-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.16
2-amino-6-chloropurine-ribose triphosphate
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
-
0.15
3'-deoxy-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.45
3'-O-Methyl-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.1557
3-[(5Z)-5-(2,3-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.0026
3-[(5Z)-5-(2,4-dichlorobenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.0073
3-[(5Z)-5-(4-tert-butylbenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]propanoic acid
0.43
6-methylthio-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
-
0.22
6-thio-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
1.5
8-bromo-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.42
8-iodoGTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
-
0.1
GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.34
ITP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.08
N1-methyl-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.15
O6-methyl-GTP
Paramecium bursaria Chlorella virus 1
-
IC50 value for formation of enzyme-GMP-complex, pH 7.5, 30°C
0.0088
[(5Z)-4-oxo-5-(4-phenoxybenzylidene)-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
0.0069
[(5Z)-5-(3,5-diiodo-2-methoxybenzylidene)-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
0.0078
[(5Z)-5-[4-(benzyloxy)benzylidene]-4-oxo-2-thioxo-1,3-thiazolidin-3-yl]acetic acid
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00003
0.000064
-
purified enzyme
0.000066
-
-
0.00007
-
purified enzyme
0.00027
-
-
0.0042
-
purified mutant, residues 1-545, D1R domain mutant, ATPase activity
0.00723
-
purified enzyme
0.03
-
purified enzyme
2.58
-
purified recombinant enzyme
7.27
-
purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7.5
-
in 50 mM Tris-HCl
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9.5
15% of maximal activity at pH 5.5, 45% at pH 9.5
6.4 - 7.9
-
pH 6.4: about 30% of activity maximum, pH 7.9: about 25% of activity maximum
6.5 - 8.5
-
pH 6.5: about 45% of activity maximum, pH 8.5: about 15% of activity maximum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
bluetongue virus serotype 10
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
-
single recombinant LEF-4 subunit is cytosolic, but the whole RNA polymerase complex is targeted to the nucleus in baculovirus infected cells
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26400
-
x * 95000, subunit containing the active site + x * 26400, transguanylyltransferase subunit, SDS-PAGE
28000
catalytically active residues 211-597, i.e. RNA guanylyltransferase domain, glycerol gradient sedimentation
31000
-
x * 95000 + x * 31000, SDS-PAGE
39000
-
x * 45000, alpha + x * 39000, beta, SDS-PAGE, probably alpha2beta2
45000
-
x * 45000, alpha + x * 39000, beta, SDS-PAGE, probably alpha2beta2
46000
residues 1-210, i.e. RNA triphosphatase domain, glycerol gradient sedimentation
48500
-
sucrose density gradient sedimentation
52000
-
x * 80000, beta, RNA 5-triphosphatase + x * 52000, alpha, mRNA guanylyltransferase activity, SDS-PAGE
54000
-
LEF-4 subunit at 0.4 M KCl, gel filtration
59000
-
x * 59000, guanylyltransferase lacking 7-methyltransferase activity, SDS-PAGE
67000
-
x * 67000, covalent enzyme-GMP complex, SDS-PAGE
68000
mRNA capping enzyme, glycerol density gradient sedimentation
69000
-
x * 69000, SDS-PAGE
80000
-
x * 80000, beta, RNA 5-triphosphatase + x * 52000, alpha, mRNA guanylyltransferase activity, SDS-PAGE
114200
-
LEF-4 subunit forming dimers in solution by weak ionic interactions, gel filtration
120000
-
copurifies with S-adenosylmethionine mRNA (guanine-7)-methyltransferase, sucrose density gradient centrifugation and gel filtration
127000
-
gel filtration, sucrose density gradient sedimentation
130000
-
gel filtration
140000
-
glycerol density gradient sedimentation
180000
-
glycerol density gradient sedimentation
additional information
-
the capping enzyme has 2 subunits: MW 95000 and 31000, the 95000 MW subunit of the vaccinia virus capping enzyme has guanylyltransferase activity, glycerol gradient centrifugation, the isolated 95000 MW guanylyltransferase can be converted to an active 60000 MW form in vitro by limited proteolysis with trypsin, the guanylyltransferase domain is localized to the amino two-thirds of the 95000 MW polypeptide
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
oligomer
-
x * 45000, alpha + x * 39000, beta, SDS-PAGE, probably alpha2beta2
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
catalytically active native MimiCE-(1-237) protein by vapor diffusion against a precipitant solution containing PEG4000 plus citrate and acetate buffers, X-ray diffraction structure determination and analysis at 1.65-2.9 A resolution
structure of residues 229-567, comprising the minimum enzymatically active human guanylyltransferase domain, to 3.0 A
-
crystals of RNA guanylyltransferase, i.e. capping enzyme, complexed with a mRNA cap analogue G(5')ppp(5')G, enzyme solution: 15 mg/ml, 1.3 mM GpppG, 50 mM Tris, pH 7.5, 0.4 M NaCl, 2 mM EDTA, 4 mM DTT, mixed with equal volume of equilibration solution: potassium phosphate 50 mM, pH 6.5, 5-10% PEG 8000, 2 mM ZnCl2, 24 h, X-ray structure determination and analysis
-
empirical and thermodynamic integration pKa estimates, along with conventional molecular dynamics simulations based on PDB entries 1ckm and 1ckn. Magnesium binding likely activates the lysine nucleophile by increasing its acidity and by biasing the deprotonated nucleophile into conformations conducive to intermediate formation
-
GTase in complex with C-terminal domain repeats of RNA polymerase Pol2 reveals a unique docking site on the nucleotidyl transferase domain for an 8-amino-acid Pol2 C-terminal domain segment, S5PPSYSPTS5P, bracketed by two Ser5-PO4 marks. At least one of the two Ser5-PO4-binding sites is required for cell viability and each site is important for cell growth at 37°C. GTase binds the Spt5 C-terminal domain at a separate docking site in the OB-fold domain that captures the Trp4 residue of the Spt5 nonapeptide repeat T1PAW4NSGSK
structures of the complete heterodimer formed by polypeptides D1 and D12. The D1 subunit comprises an N-terminal RNA triphosphatase (TPase)-guanylyltransferase (GTase) module and a C-terminal guanine-N7-methyltransferase (MTase) module. The D12 subunit binds and allosterically stimulates the MTase module. An extensive TPase-GTase interface clamps the GTase nucleotidyltransferase and oligosaccharide-binding fold domains in a closed conformation around GTP
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7
-
the amount of G(5')ppp(5')RNA formed at pH 5.5 is 50% of the amount formed at pH 7
674811
additional information
bluetongue virus serotype 10
-
enzyme-GMP complex is resistant to alkali, but is cleaved at acidic conditions with HCl or NH2OH
643489
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
5 min, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
can be frozen and thawed several times without apparent loss of activity
-
stable after a few cycles of freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 10 mM Tris-HCl buffer, pH 8, 10 mM 2-mercaptoethanol, 50% glycerol, 0.1 mg/ml gelatin, enzyme concentration: 0.1 mg/ml, stable
-
-20°C, stable for more than 6 months, considerably longer at -70°C
-
-70°C, stable for at least 9 months
-
-80°C, reaction intermediate enzyme-GMP, purified, stable for at least several weeks
-
-80°C, stable for at least 6 months
-
0°C, 48 h, 90% loss of activity, 59000 MW protein which lacks 7-methyltransferase activity
-
4°C or -20°C, purified enzyme, stable for several months
-
4°C, stable for more than 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1000fold, recombinant from Escherichia coli
-
231fold; contains little or no RNA 5'-triphosphatase or methyltransferase activity
-
800fold; partial; physically associated with mRNA 5'-triphosphatase activity
-
copurifies with S-adenosylmethionine mRNA (guanine-7)-methyltransferase
-
large scale, to homogeneity, enzyme has mRNA guanylyltransferase activity
-
mutant enzyme C-terminally truncated to residues 1-545
-
nickel-agarose chromatography
-
physically associated with mRNA 5'-triphosphatase activity
-
physically associated with mRNA 5'-triphosphatase activity; to homogeneity
-
recombinant from Spodoptera frugiperda cells after infection with baculovirus, to homogeneity
bluetongue virus serotype 10
-
recombinant from vaccinia virus grown in HeLa cells
-
recombinant His-tagged full length enzyme as well as the recombinant His-tagged N-terminal and C-terminal domains from Escherichia coli BL21(DE3)
recombinant His-tagged wild-type and mutant K177A protein from Bacillus megaterium
-
recombinant His6-tagged A103R protein from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant LEF-4 subunit from overexpressing Spodoptera frugiperda cells
-
recombinant protein
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recombinant truncated mutant, residues 438-597, His-tagged
recombinant wild-type and mutant K226A from Trichoplusia ni or Spodoptera frugiperda SF21 cells via baculovirus infection
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis
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DNA and amino acid sequence determination and analysis, expression of truncation mutant in Escherichia coli BL21(DE3) and wild-type and mutant K294A enzyme in Saccharomyces haploid deficient mutant strain, functional complementation of the latter by wild-type enzyme; in vitro translation of wild-type and mutant enzyme
expession in Escherichia coli
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expressed in Escherichia coli BL21(ROS2) cells
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expression in Escherichia coli
expression in HeLa cells via vaccinia virus strain WR into whose thymidine kinase gene the reovirus lambda2 genome segment has been inserted, possesses neither nucleoside nor RNA 5'-triphosphatase activity nor methyltransferase activity
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expression in Saccharomyces cerevisiae
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expression in Spodoptera frugiperda cells via infection with baculovirus
bluetongue virus serotype 10
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expression of His-tagged full length enzyme, residues 211-597, comprising the catalytical domain, and residues 1-210 in Escherichia coli BL21(DE3); expression of wild-type full length enzyme and catalytic domain in deficient Saccharomyces cerevisiae strain YBS2 as His-tagged protein, functional complementation by both of them; mRNA capping enzyme, DNA sequence determination and analysis
expression of viral D1 and D12 orfs encoded subunits in Escherichia coli BL21(DE3)
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expression of wild-type and mutant K177A in Bacillus megaterium as His-tagged proteins
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expression of wild-type from plasmid in deficient mutant ceg1-25
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gene A103R, expression of His6-tagged A103R protein in Escherichia coli strain BL21(DE3)
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gene CEG1 from genomic library, expression of catalytically active alpha-subunit in Escherichia coli, chromosome mapping
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gene CEG1, guanylylpeptide, DNA and amino acid sequence determination
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gene L2, expression of wild-type and mutant K226A protein lambda2 in Trichoplusia ni or Spodoptera frugiperda SF21 cells via baculovirus infection
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mutant lacking the C-terminal domain, consisting of the D1R domain, residues 1-545, expression in Escherichia coli BL21(DE3)pLysS
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overexpression of LEF-4 subunit in Spodoptera frugiperda cells by recombinant virus infection under control of polyhedrin promotor
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RNA genome sequence determination and analysis, expression of lambdaC in Spodoptera frugiperda cells via baculovirus infection
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D468A
site-directed mutagenesis, inactive mutant
K292A
site-directed mutagenesis, inactive mutant
K496A
site-directed mutagenesis, inactive mutant
K498A
site-directed mutagenesis, inactive mutant
H68A
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mutation enhances GTP methylation reaction but disables the following transguanylation reaction
H68C
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mutation results in a change in the nature nature of the bond linking the enzyme and m7GMP, suggesting that residue H68 covalently binds to m7GMP in the intermediate
D532A
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inactive
E234A
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mutation does not inhibit the formation of the phosphoamide intermediate
K458A
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inactive
K460A
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inactive
K533A
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inactive
R528A
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poor activity
R530A
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inactive
K177A
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site-directed mutagenesis, no activity
K171A
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site-directed mutagenesis, 0.22% of wild-type autoguanylylation activity
K190A
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site-directed mutagenesis, autoguanylylation inactive
K197A
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site-directed mutagenesis, 7.5% of wild-type autoguanylylation activity
K226A
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site-directed mutagenesis, unaltered properties
K44A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
K89A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
K94A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
K171A
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site-directed mutagenesis, 0.22% of wild-type autoguanylylation activity
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K190A
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site-directed mutagenesis, autoguanylylation inactive
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K44A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
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K89A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
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K94A
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site-directed mutagenesis, approximately wild-type autoguanylylation activity levels
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K175A
no loss of interaction with polymerase II C-terminal domain
K198A
no loss of interaction with polymerase II C-terminal domain
R159A
loss of interaction with polymerase II C-terminal domain. Residue Arg159 of Ceg1 interacts strongly with polymerase II C-terminal domain
R185A
loss of interaction with polymerase II C-terminal domain. Residue Arg185 of Ceg1 interacts strongly with polymerase II C-terminal domain
G164T
56ld increase in Kd value for Pol2 CTD–Ser5-PO4
G164T/H201N
349foldd increase in Kd value for Pol2 CTD–Ser5-PO4
H201N
46ld increase in Kd value for Pol2 CTD–Ser5-PO4
H201N/R364A/Y368F
98fold increase in Kd value for Pol2 CTD–Ser5-PO4
R157E
85fold increase in Kd value for Pol2 CTD–Ser5-PO4
R157E/H201N
526fold increase in Kd value for Pol2 CTD–Ser5-PO4
G164T
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56ld increase in Kd value for Pol2 CTD–Ser5-PO4
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G164T/H201N
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349foldd increase in Kd value for Pol2 CTD–Ser5-PO4
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H201N
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46ld increase in Kd value for Pol2 CTD–Ser5-PO4
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R157E
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85fold increase in Kd value for Pol2 CTD–Ser5-PO4
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R157E/H201N
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526fold increase in Kd value for Pol2 CTD–Ser5-PO4
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E192A
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113% of wild-type guanylyltransferase activity, 4% of wild-type RNA triphosphatase activity
K478A
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1% of wild-type guanylyltransferase activity, 42% of wild-type RNA triphosphatase activity
L47A/L50A/T51A
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1% of wild-type guanylyltransferase activity, 43% of wild-type RNA triphosphatase activity
N181A
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48% of wild-type guanylyltransferase activity, 69% of wild-type RNA triphosphatase activity
R186A
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51% of wild-type guanylyltransferase activity, 92% of wild-type RNA triphosphatase activity
T10A
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19% of wild-type guanylyltransferase activity, 104% of wild-type RNA triphosphatase activity
E192A
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113% of wild-type guanylyltransferase activity, 4% of wild-type RNA triphosphatase activity
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K478A
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1% of wild-type guanylyltransferase activity, 42% of wild-type RNA triphosphatase activity
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N181A
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48% of wild-type guanylyltransferase activity, 69% of wild-type RNA triphosphatase activity
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R186A
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51% of wild-type guanylyltransferase activity, 92% of wild-type RNA triphosphatase activity
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T10A
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19% of wild-type guanylyltransferase activity, 104% of wild-type RNA triphosphatase activity
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additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
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enzyme can be used as a tool for specific 5'-end-labeling of mRNA
Show AA Sequence (766 entries)
Please use the Sequence Search for a specific query.