BRENDA - Enzyme Database
show all sequences of 2.1.1.57

Recognition of capped RNA substrates by VP39, the vaccinia virus-encoded mRNA cap-specific 2'-O-methyltransferase

Lockless, S.W.; Cheng, H.T.; Hodel, A.E.; Quiocho, F.A.; Gershon, P.D.; Biochemistry 37, 8564-8574 (1998)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
mutant enzymes are expressed in Escherichia coli and the tag is cleaved off
Vaccinia virus
Engineering
Protein Variants
Commentary
Organism
D182A
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
D182A/E233A
m7G binding pocket double mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
E233A
m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
F180A
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
Y22A
m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Vaccinia virus
0.000005
0.000015
RNA chain length 20-50 nt
-
Vaccinia virus
0.00016
0.00023
RNA chain length 2-6 nt
-
Vaccinia virus
Organism
Organism
UniProt
Commentary
Textmining
Vaccinia virus
-
VP39 protein
-
Purification (Commentary)
Purification (Commentary)
Organism
mutants recombinant from E. coli
Vaccinia virus
Reaction
Reaction
Commentary
Organism
Reaction ID
S-adenosyl-L-methionine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(purine-ribonucleotide)-[mRNA] = S-adenosyl-L-homocysteine + a 5'-(N7-methyl 5'-triphosphoguanosine)-(2'-O-methyl-purine-ribonucleotide)-[mRNA]
binding sites of the enzyme for the terminal cap nucleotide, m7G, the cap-proximal RNA, and for RNA, immediately downstream, are characterized
Vaccinia virus
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 + ApppGR-RNA
adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + ApppGR-RNA
synthetic substrate with unusual cap structure, 20% activity compared to m7G-terminated substrates
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + m7G(5')pppR-RNA
adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7G(5')pppRm-RNA
-
-
-
?
S-adenosyl-L-methionine + m7GpppGp
substrate of minimal chain length for the cap-specific enzyme, m7GpppG is no substrate
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7GpppGmp
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpN
-
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7GpppGmpN
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpNp
-
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpUbiotin-p
-
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + RNA chain length 2-6 nt
-
639484
Vaccinia virus
?
-
-
-
r
S-adenosyl-L-methionine + RNA chain length 20-50 nt
-
639484
Vaccinia virus
?
-
-
-
r
Synonyms
Synonyms
Commentary
Organism
mRNA cap-specific 2'-O-methyltransferase
i.e. VP39
Vaccinia virus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Vaccinia virus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6
-
assay at
Vaccinia virus
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.5
8
m7G- and A-capped RNA substrates
Vaccinia virus
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
additional information
-
binding site of the enzyme for RNA, immediately downstream, is pH-sensitive, above pH 7.5: dramatic increase of Km
Vaccinia virus
Cloned(Commentary) (protein specific)
Commentary
Organism
mutant enzymes are expressed in Escherichia coli and the tag is cleaved off
Vaccinia virus
Engineering (protein specific)
Protein Variants
Commentary
Organism
D182A
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
D182A/E233A
m7G binding pocket double mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
E233A
m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
F180A
m7G binding pocket mutant, mutagenesis, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
Y22A
m7G binding pocket mutant, mutagenesis, N-terminal glutathione-S-transferase-tagged, C-terminal truncated by 26 amino acids, lacks specific m7G-contact side chains and shows reduced activity, but remains cap-dependent
Vaccinia virus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Vaccinia virus
0.000005
0.000015
RNA chain length 20-50 nt
-
Vaccinia virus
0.00016
0.00023
RNA chain length 2-6 nt
-
Vaccinia virus
Purification (Commentary) (protein specific)
Commentary
Organism
mutants recombinant from E. coli
Vaccinia virus
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 + ApppGR-RNA
adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + ApppGR-RNA
synthetic substrate with unusual cap structure, 20% activity compared to m7G-terminated substrates
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + m7G(5')pppR-RNA
adenine-capped RNA, wild-type and mutants, indicating the redundancy of m7G-contact residues able to confer cap-type specificity
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7G(5')pppRm-RNA
-
-
-
?
S-adenosyl-L-methionine + m7GpppGp
substrate of minimal chain length for the cap-specific enzyme, m7GpppG is no substrate
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7GpppGmp
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpN
-
639484
Vaccinia virus
S-adenosyl-L-homocysteine + m7GpppGmpN
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpNp
-
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + m7GpppGpUbiotin-p
-
639484
Vaccinia virus
?
-
-
-
?
S-adenosyl-L-methionine + RNA chain length 2-6 nt
-
639484
Vaccinia virus
?
-
-
-
r
S-adenosyl-L-methionine + RNA chain length 20-50 nt
-
639484
Vaccinia virus
?
-
-
-
r
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Vaccinia virus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6
-
assay at
Vaccinia virus
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.5
8
m7G- and A-capped RNA substrates
Vaccinia virus
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
additional information
-
binding site of the enzyme for RNA, immediately downstream, is pH-sensitive, above pH 7.5: dramatic increase of Km
Vaccinia virus
Other publictions for EC 2.1.1.57
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)
757292
Simabuco
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5597-5611
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757922
Toczydlowska-Socha
Human RNA cap1 methyltransfer ...
Homo sapiens
Philos. Trans. R. Soc. Lond. B Biol. Sci.
373
20180161
2018
1
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1
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1
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4
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1
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1
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758536
Quirin
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1
1
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733878
Stewart
Structure-based identification ...
bluetongue virus
FEBS open bio
5
138-146
2015
-
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1
1
6
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1
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4
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6
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1
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1
1
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720881
Szretter
2-O methylation of the viral m ...
West Nile virus
PLoS Pathog.
8
e1002698
2012
-
-
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1
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5
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-
1
1
-
-
-
721104
Ke
Short peptides derived from th ...
Severe acute respiratory syndrome-related coronavirus, Severe acute respiratory syndrome-related coronavirus WHU
Virus Res.
167
322-328
2012
1
-
1
-
-
-
1
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5
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1
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2
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5
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1
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1
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1
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720878
Decroly
Crystal structure and function ...
Severe acute respiratory syndrome-related coronavirus
PLoS Pathog.
7
e1002059
2011
1
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31
-
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1
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4
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31
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1
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720879
Chen
Biochemical and structural ins ...
Severe acute respiratory syndrome-related coronavirus, Severe acute respiratory syndrome-related coronavirus WHU
PLoS Pathog.
7
e1002294
2011
1
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1
1
4
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1
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6
-
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1
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4
1
4
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1
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1
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1
4
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1
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4
1
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704920
Selisko
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Dengue virus, feline coronavirus
J. Gen. Virol.
91
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2010
8
-
2
-
11
-
11
2
-
2
2
6
-
7
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2
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6
1
10
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2
1
2
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3
8
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2
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11
-
3
11
-
2
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2
2
6
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2
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6
1
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-
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2
1
2
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719891
Belanger
Characterization of hMTr1, a h ...
Homo sapiens
J. Biol. Chem.
285
33037-33044
2010
-
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1
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1
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2
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1
1
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5
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1
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1
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5
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1
1
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1
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1
1
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1
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719892
Lugari
Molecular mapping of the RNA c ...
Severe acute respiratory syndrome-related coronavirus
J. Biol. Chem.
285
33230-33241
2010
1
1
1
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15
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5
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1
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15
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721099
Chung
Higher catalytic efficiency of ...
Dengue virus
Virology
402
52-60
2010
-
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1
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2
2
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6
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1
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1
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3
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1
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721101
Zhang
Identification of aromatic ami ...
vesicular stomatitis virus
Virology
408
241-252
2010
-
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10
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1
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1
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2
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4
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10
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1
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2
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671151
Mastrangelo
Preliminary characterization o ...
Meaban virus, Yokose virus
Acta Crystallogr. Sect. F
F62
768-770
2006
-
2
2
2
-
-
-
-
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8
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2
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4
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2
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2
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2
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-
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-
-
-
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-
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-
-
-
-
672205
Li
pKa of the mRNA cap-specific 2 ...
Vaccinia virus
Biochemistry
45
907-917
2006
-
1
1
-
2
-
-
-
-
-
-
-
-
2
-
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-
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1
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2
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1
1
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2
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1
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677099
Arhin
A protein related to the vacci ...
Trypanosoma brucei
RNA
12
53-62
2006
-
2
-
-
2
-
-
-
1
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2
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9
-
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5
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2
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2
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1
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675702
Li
Amino acid residues within con ...
vesicular stomatitis virus
J. Virol.
79
13373-13384
2005
-
1
1
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3
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4
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1
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6
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