BRENDA - Enzyme Database
show all sequences of 2.1.1.56

Structure and mechanism of mRNA cap (guanine-N7) methyltransferase

Fabrega, C.; Hausmann, S.; Shen, V.; Shuman, S.; Lima, C.D.; Mol. Cell 13, 77-89 (2004)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expression of N-terminally His6-tagged enzyme, residues 1-298, as Smt3-fusion protein in Escherichia coli strain BL21(DE3) or strain B834DE3, expression of wild-type and mutant alleles under control of a yeast promotor in a Saccharomyces cerevisiae abd1DELTA null mutant strain
Encephalitozoon cuniculi
Crystallization (Commentary)
Crystallization (Commentary)
Organism
6.5 mg/ml purified native or selenomethionine-labeled, or Hg-labeled enzyme in complex with S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, or the cap guanylate plus S-adenosyl-L-homocysteine, in 20 mM Tris-HCl, pH 8.0, 220 mM NaCl, vapour diffusion against a well solution of 1.2 M sodium potassium tartrate, 50 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, pH 6.0-6.2, and 20 mM DTT, X-ray diffraction structure determination and analysis at 2.0-2.7 A resolution, modeling
Encephalitozoon cuniculi
Engineering
Protein Variants
Commentary
Organism
D122A/S123A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, only slightly at 37C
Encephalitozoon cuniculi
D78A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant at 30C and 37C, but at 25C
Encephalitozoon cuniculi
D94A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
F141A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
I95A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C, 30C, and 37C, no effect on enzyme function
Encephalitozoon cuniculi
I95A/Y124A
site-directed mutagenesis, only slight complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, growth defects
Encephalitozoon cuniculi
K54A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, inactive mutant in vivo
Encephalitozoon cuniculi
K75A
site-directed mutagenesis, some complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, temperature-sensitive phenotype
Encephalitozoon cuniculi
K81A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
L216A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, no effect on cell growth
Encephalitozoon cuniculi
N50A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, only modest growth defects
Encephalitozoon cuniculi
N50A/Y284A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
N51A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C
Encephalitozoon cuniculi
R106A
site-directed mutagenesis, only slight complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C
Encephalitozoon cuniculi
R47A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C
Encephalitozoon cuniculi
R47A/K75A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
Y124A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, temperature-sensitive phenotype
Encephalitozoon cuniculi
Y284A
site-directed mutagenesis, some complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, slow growth phenotype
Encephalitozoon cuniculi
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
S-adenosyl-L-methionine + G(5')pppR-RNA
Encephalitozoon cuniculi
essential and defining step in the eukaryotic mRNA synthesis
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Encephalitozoon cuniculi
-
microsporidian parasite, enzyme Ecm1
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant N-terminally His6-tagged enzyme as Smt3-fusion protein from Escherichia coli strain BL21(DE3), removal of His-Smt3 tag by specific protease Ulp1, purification by metal affinity chromatography and gel filtration
Encephalitozoon cuniculi
Reaction
Reaction
Commentary
Organism
Reaction ID
S-adenosyl-L-methionine + a 5'-(5'-triphosphoguanosine)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N7-methyl 5'-triphosphoguanosine)-[mRNA]
in-line mechanism of methyl transfer without direct contact between the enzyme and the N7 atom of guanine, which is the attacking nucleophile, the methyl carbon of S-adenosyl-L-methionine, or the sulfur of S-adenosyl-L-methionine/S-adenosyl-L-homocysteine, enzyme-ligand structures, active site structure
Encephalitozoon cuniculi
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
S-adenosyl-L-methionine + G(5')pppR-RNA
essential and defining step in the eukaryotic mRNA synthesis
659921
Encephalitozoon cuniculi
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
-
?
S-adenosyl-L-methionine + G(5')pppR-RNA
enzyme contains the conserved S-adenosyl-L-methionine-binding motif VLDLGXGXG, structural features for cap guanine specificity
659921
Encephalitozoon cuniculi
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
-
?
Subunits
Subunits
Commentary
Organism
More
primary sequence, sequence comparison with other species, enzyme-ligand structures
Encephalitozoon cuniculi
Synonyms
Synonyms
Commentary
Organism
mRNA cap (guanine-N7) methyltransferase
-
Encephalitozoon cuniculi
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of N-terminally His6-tagged enzyme, residues 1-298, as Smt3-fusion protein in Escherichia coli strain BL21(DE3) or strain B834DE3, expression of wild-type and mutant alleles under control of a yeast promotor in a Saccharomyces cerevisiae abd1DELTA null mutant strain
Encephalitozoon cuniculi
Crystallization (Commentary) (protein specific)
Crystallization
Organism
6.5 mg/ml purified native or selenomethionine-labeled, or Hg-labeled enzyme in complex with S-adenosyl-L-methionine, S-adenosyl-L-homocysteine, or the cap guanylate plus S-adenosyl-L-homocysteine, in 20 mM Tris-HCl, pH 8.0, 220 mM NaCl, vapour diffusion against a well solution of 1.2 M sodium potassium tartrate, 50 mM bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane, pH 6.0-6.2, and 20 mM DTT, X-ray diffraction structure determination and analysis at 2.0-2.7 A resolution, modeling
Encephalitozoon cuniculi
Engineering (protein specific)
Protein Variants
Commentary
Organism
D122A/S123A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, only slightly at 37C
Encephalitozoon cuniculi
D78A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant at 30C and 37C, but at 25C
Encephalitozoon cuniculi
D94A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
F141A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
I95A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C, 30C, and 37C, no effect on enzyme function
Encephalitozoon cuniculi
I95A/Y124A
site-directed mutagenesis, only slight complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, growth defects
Encephalitozoon cuniculi
K54A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, inactive mutant in vivo
Encephalitozoon cuniculi
K75A
site-directed mutagenesis, some complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, temperature-sensitive phenotype
Encephalitozoon cuniculi
K81A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
L216A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, no effect on cell growth
Encephalitozoon cuniculi
N50A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C, only modest growth defects
Encephalitozoon cuniculi
N50A/Y284A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
N51A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C
Encephalitozoon cuniculi
R106A
site-directed mutagenesis, only slight complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C
Encephalitozoon cuniculi
R47A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, slightly reduced at 37C
Encephalitozoon cuniculi
R47A/K75A
site-directed mutagenesis, no complementation of the yeast abd1DELTA mutant, mutation is lethal in vivo
Encephalitozoon cuniculi
Y124A
site-directed mutagenesis, complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, temperature-sensitive phenotype
Encephalitozoon cuniculi
Y284A
site-directed mutagenesis, some complementation of the yeast abd1DELTA mutant at 25C and 30C, not at 37C, slow growth phenotype
Encephalitozoon cuniculi
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
S-adenosyl-L-methionine + G(5')pppR-RNA
Encephalitozoon cuniculi
essential and defining step in the eukaryotic mRNA synthesis
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His6-tagged enzyme as Smt3-fusion protein from Escherichia coli strain BL21(DE3), removal of His-Smt3 tag by specific protease Ulp1, purification by metal affinity chromatography and gel filtration
Encephalitozoon cuniculi
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
S-adenosyl-L-methionine + G(5')pppR-RNA
essential and defining step in the eukaryotic mRNA synthesis
659921
Encephalitozoon cuniculi
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
-
?
S-adenosyl-L-methionine + G(5')pppR-RNA
enzyme contains the conserved S-adenosyl-L-methionine-binding motif VLDLGXGXG, structural features for cap guanine specificity
659921
Encephalitozoon cuniculi
S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
primary sequence, sequence comparison with other species, enzyme-ligand structures
Encephalitozoon cuniculi
Other publictions for EC 2.1.1.56
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733122
Sun
Yeast-based assays for the hig ...
SARS coronavirus
Antiviral Res.
104
156-164
2014
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1
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1
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734586
Chen
Structure-function analysis of ...
SARS coronavirus
J. Virol.
87
6296-6305
2013
-
-
1
1
27
-
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-
-
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-
4
-
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1
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1
27
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735335
Jin
Characterization of the guanin ...
SARS coronavirus
Virus Res.
176
45-52
2013
-
-
-
-
18
-
-
-
-
-
-
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-
5
-
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6
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18
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6
-
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-
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-
-
-
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-
735240
Thillier
Synthesis of 5' cap-0 and cap- ...
Homo sapiens
RNA
18
856-868
2012
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
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-
-
-
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1
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1
1
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-
704920
Selisko
Biochemical characterization o ...
Dengue virus
J. Gen. Virol.
91
112-121
2010
-
-
-
-
-
-
1
-
-
-
-
1
-
3
-
-
-
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-
-
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-
1
-
4
-
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-
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1
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1
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1
-
-
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-
-
-
-
-
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-
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-
721099
Chung
Higher catalytic efficiency of ...
Dengue virus
Virology
402
52-60
2010
-
-
-
-
-
-
2
2
-
-
-
-
-
1
-
-
-
-
-
-
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1
-
1
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-
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2
-
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-
2
-
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-
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-
-
2
2
-
2
-
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1
-
-
-
-
2
-
-
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-
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-
-
-
721100
Dong
Biochemical and genetic charac ...
Dengue virus
Virology
405
568-578
2010
-
-
1
-
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
2
-
2
-
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1
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1
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4
-
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1
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-
-
2
-
-
-
-
-
1
-
-
-
-
1
1
-
-
-
721101
Zhang
Identification of aromatic ami ...
vesicular stomatitis virus
Virology
408
241-252
2010
-
-
-
-
16
-
-
-
-
-
-
-
-
1
-
-
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-
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1
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1
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1
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1
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16
-
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1
-
-
-
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701751
Milani
Flaviviral methyltransferase/R ...
Dengue virus, Wesselsbron virus
Antiviral Res.
83
28-34
2009
-
-
-
-
-
-
4
-
-
-
-
-
-
4
-
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2
-
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2
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6
-
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2
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2
4
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2
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2
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706020
Cowling
Enhanced mRNA cap methylation ...
Homo sapiens
Oncogene
29
930-936
2009
-
-
-
-
-
-
-
-
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-
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1
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2
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1
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3
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2
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1
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1
1
-
-
-
706535
Chen
Functional screen reveals SARS ...
SARS coronavirus
Proc. Natl. Acad. Sci. USA
106
3484-3489
2009
-
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1
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4
-
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6
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1
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4
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1
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4
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1
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690028
Zheng
Mutational analysis of vaccini ...
Vaccinia virus
RNA
14
2297-2304
2008
-
-
-
-
20
-
-
-
-
-
-
-
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2
-
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1
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1
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20
-
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1
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705423
Dong
West Nile virus methyltransfer ...
West Nile virus
J. Virol.
82
4295-4307
2008
-
-
-
-
20
-
1
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-
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-
1
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1
-
3
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-
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20
-
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1
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1
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-
-
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-
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1
1
-
-
-
706721
Zheng
Structure-function analysis of ...
Vaccinia virus
RNA
14
696-705
2008
-
-
1
-
49
-
-
16
-
-
-
-
-
2
-
-
1
-
-
-
-
-
1
1
2
-
-
-
14
-
-
-
-
-
-
-
-
-
1
-
-
49
-
-
-
-
16
-
-
-
-
-
-
-
1
-
-
-
-
1
1
-
-
-
14
-
-
-
-
-
-
-
-
-
-
676231
Peyrane
High-yield production of short ...
Homo sapiens
Nucleic Acids Res.
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Identification of a unique gua ...
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Pugh
Effects of S-adenosylhomocyste ...
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5 Cap methylation of homologou ...
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Purification of mRNA guanylylt ...
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Pugh
Inhibition of Newcastle diseas ...
Avian avulavirus 1
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Ensinger
Modification of the 5 terminus ...
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mRNA guanylyltransferase and m ...
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Ensinger
Modification of the 5-terminus ...
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72
2525-2529
1975
-
-
-
-
-
-
2
-
1
2
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
1
2
-
-
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-