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Literature summary for 2.1.1.57 extracted from

  • Selisko, B.; Peyrane, F.F.; Canard, B.; Alvarez, K.; Decroly, E.
    Biochemical characterization of the (nucleoside-2'O)-methyltransferase activity of dengue virus protein NS5 using purified capped RNA oligonucleotides (7Me)GpppAC(n) and GpppAC(n) (2010), J. Gen. Virol., 91, 112-121.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
2',3',5'-tri-O-acetyladenosine about 20% increased activity at 0.1 mM Dengue virus
2',3',5'-tri-O-acetyladenosine about 50% increased activity at 0.1 mM feline coronavirus
3-deazaadenosine about 10% increased activity at 0.1 mM feline coronavirus
5'-deoxy-5'-(methylthio)adenosine about 75% increased activity at 0.1 mM feline coronavirus
5'-deoxy-5'-methylthioadenosine about 10% increased activity at 0.1 mM Dengue virus
5'-S-isobutylthio-5'-deoxyadenosine about 60% increased activity at 0.1 mM feline coronavirus
ribavirin about 10% increased activity at 0.1 mM feline coronavirus
ribavirin triphosphate about 20% increased activity at 0.1 mM feline coronavirus

Cloned(Commentary)

Cloned (Comment) Organism
-
Dengue virus
expressed in Escherichia coli C41(DE3) cells feline coronavirus

Protein Variants

Protein Variants Comment Organism
D113A about 110% substrate binding and activity compared to the wild type enzyme feline coronavirus
D129A about 40% substrate binding and activity compared to the wild type enzyme feline coronavirus
D221A about 115% substrate binding and activity compared to the wild type enzyme feline coronavirus
D247A about 75% substrate binding and activity compared to the wild type enzyme feline coronavirus
E202A about 30% substrate binding and activity compared to the wild type enzyme feline coronavirus
F173A about 20% substrate binding and activity compared to the wild type enzyme feline coronavirus
K169A about 20% substrate binding and activity compared to the wild type enzyme feline coronavirus
K45A about 1% substrate binding and activity compared to the wild type enzyme feline coronavirus
W4A about 90% substrate binding and activity compared to the wild type enzyme feline coronavirus
Y14A about 10% substrate binding and activity compared to the wild type enzyme feline coronavirus
Y29A about 15% substrate binding and activity compared to the wild type enzyme feline coronavirus

Inhibitors

Inhibitors Comment Organism Structure
5'-S-isobutylthio-5'-deoxyadenosine about 25% inhibition at 0.1 mM Dengue virus
5-ethynyl-1-beta-D-ribofuranosylimidazole-4-carboxamide triphosphate about 40% inhibition at 0.1 mM Dengue virus
deazaadenosine about 5% inhibition at 0.1 mM Dengue virus
ribavirin about 10% inhibition at 0.1 mM Dengue virus
ribavirin triphosphate about 50% inhibition at 0.1 mM Dengue virus
S-(5'-adenosyl)-L-cysteine about 25% inhibition at 0.1 mM feline coronavirus
S-5'-adenosyl-L-cysteine about 5% inhibition at 0.1 mM Dengue virus
S-adenosyl-L-homocysteine complete inhibition at 0.1 mM Dengue virus
S-adenosyl-L-homocysteine about 25% inhibition at 0.1 mM feline coronavirus
sinefungin complete inhibition at 0.1 mM Dengue virus
sinefungin about 5% inhibition at 0.1 mM feline coronavirus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000394
-
7Me-GpppA-C5 in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus
0.00326
-
S-adenosyl-L-methionine in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus

Metals/Ions

Metals/Ions Comment Organism Structure
K+ 110% activity in the presence of 10 mM KCl, in Tris-HCl buffer (pH 7.5) Dengue virus
Mg2+ maximum activity in the presence of 1 mM feline coronavirus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
2 * 35000, SDS-PAGE feline coronavirus
65000
-
gel filtration feline coronavirus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Dengue virus maximum 2'O-MTase activity requires the cap and at least 6 nucleotides ?
-
?
additional information feline coronavirus the enzyme shows highest activity towards 7Me-GpppA-Cn where n equals at least four, and does not methylate GpppAC5 or pppAC5 ?
-
?
S-adenosyl-L-methionine + 7Me-GpppA-C5 feline coronavirus
-
S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
?
S-adenosyl-L-methionine + 7Me-GpppA-C5 Dengue virus exclusive methylation at the 2'O position S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
?
S-adenosyl-L-methionine + 7Me-GpppA-Cn Dengue virus exclusive methylation at the 2'O position S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-Cn
-
?
S-adenosyl-L-methionine + GpppA-Cn Dengue virus exclusive methylation at the 2'O position S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
-
?

Organism

Organism UniProt Comment Textmining
Dengue virus
-
-
-
feline coronavirus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Dengue virus
Sepharose resin column chromatography and Superdex 200 gel filtration feline coronavirus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information maximum 2'O-MTase activity requires the cap and at least 6 nucleotides Dengue virus ?
-
?
additional information the enzyme shows highest activity towards 7Me-GpppA-Cn where n equals at least four, and does not methylate GpppAC5 or pppAC5 feline coronavirus ?
-
?
S-adenosyl-L-methionine + 7Me-GpppA-C5
-
feline coronavirus S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
?
S-adenosyl-L-methionine + 7Me-GpppA-C5 exclusive methylation at the 2'O position Dengue virus S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
?
S-adenosyl-L-methionine + 7Me-GpppA-Cn exclusive methylation at the 2'O position Dengue virus S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-Cn
-
?
S-adenosyl-L-methionine + GpppA-Cn exclusive methylation at the 2'O position Dengue virus S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 35000, SDS-PAGE feline coronavirus

Synonyms

Synonyms Comment Organism
(nucleoside-2'O)-methyltransferase part of the NS5 protein Dengue virus
2'O-MTase
-
Dengue virus
2'O-MTase
-
feline coronavirus
nonstructural protein 16
-
feline coronavirus
nonstructural protein 16 methyltransferase
-
feline coronavirus
nonstructural protein 5 methyltransferase
-
Dengue virus
NS5MTase the protein contains mRNA cap (guanine N-7) methyltransferase and (nucleoside-2'O)-methyltransferase activities Dengue virus
nsp16
-
feline coronavirus
RNA (nucleoside-2'O)-methyltransferase
-
feline coronavirus
S-adenosyl-L-methionine-dependent mRNA cap MTase
-
feline coronavirus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
feline coronavirus
8
-
-
Dengue virus

pH Range

pH Minimum pH Maximum Comment Organism
7 9
-
Dengue virus

pH Stability

pH Stability pH Stability Maximum Comment Organism
10
-
at basic pH, 2'O-MTase activity is practically non-existent and neither Mg2+ nor KCl is able to activate the methyl transfer at pH 10.0 Dengue virus
10
-
at basic pH, 2'O-MTase activity is practically non-existent and neither Mg2+ nor KCl is able to activate the methyl transfer at pH 10.0 feline coronavirus

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00034
-
in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus S-adenosyl-L-homocysteine
0.00063
-
in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus sinefungin
0.144
-
-
feline coronavirus S-adenosyl-L-homocysteine