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Literature summary 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

EC Number Activating Compound Comment Organism Structure
2.1.1.57 2',3',5'-tri-O-acetyladenosine about 20% increased activity at 0.1 mM Dengue virus
2.1.1.57 2',3',5'-tri-O-acetyladenosine about 50% increased activity at 0.1 mM feline coronavirus
2.1.1.57 3-deazaadenosine about 10% increased activity at 0.1 mM feline coronavirus
2.1.1.57 5'-deoxy-5'-(methylthio)adenosine about 75% increased activity at 0.1 mM feline coronavirus
2.1.1.57 5'-deoxy-5'-methylthioadenosine about 10% increased activity at 0.1 mM Dengue virus
2.1.1.57 5'-S-isobutylthio-5'-deoxyadenosine about 60% increased activity at 0.1 mM feline coronavirus
2.1.1.57 ribavirin about 10% increased activity at 0.1 mM feline coronavirus
2.1.1.57 ribavirin triphosphate about 20% increased activity at 0.1 mM feline coronavirus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.57
-
 Dengue virus
2.1.1.57 expressed in Escherichia coli C41(DE3) cells feline coronavirus

Protein Variants

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.56 sinefungin
-
 Dengue virus
2.1.1.57 5'-S-isobutylthio-5'-deoxyadenosine about 25% inhibition at 0.1 mM Dengue virus
2.1.1.57 5-ethynyl-1-beta-D-ribofuranosylimidazole-4-carboxamide triphosphate about 40% inhibition at 0.1 mM Dengue virus
2.1.1.57 deazaadenosine about 5% inhibition at 0.1 mM Dengue virus
2.1.1.57 ribavirin about 10% inhibition at 0.1 mM Dengue virus
2.1.1.57 ribavirin triphosphate about 50% inhibition at 0.1 mM Dengue virus
2.1.1.57 S-(5'-adenosyl)-L-cysteine about 25% inhibition at 0.1 mM feline coronavirus
2.1.1.57 S-5'-adenosyl-L-cysteine about 5% inhibition at 0.1 mM Dengue virus
2.1.1.57 S-adenosyl-L-homocysteine complete inhibition at 0.1 mM Dengue virus
2.1.1.57 S-adenosyl-L-homocysteine about 25% inhibition at 0.1 mM feline coronavirus
2.1.1.57 sinefungin complete inhibition at 0.1 mM Dengue virus
2.1.1.57 sinefungin about 5% inhibition at 0.1 mM feline coronavirus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.57 0.000394
-
 7Me-GpppA-C5 in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus
2.1.1.57 0.00326
-
 S-adenosyl-L-methionine in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.57 35000
-
 2 * 35000, SDS-PAGE feline coronavirus
2.1.1.57 65000
-
 gel filtration feline coronavirus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.56 S-adenosyl-L-methionine + G(5')pppR-RNA Dengue virus
-
 S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
 ?
2.1.1.57 additional information Dengue virus maximum 2'O-MTase activity requires the cap and at least 6 nucleotides ?
-
 ?
2.1.1.57 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 ?
-
 ?
2.1.1.57 S-adenosyl-L-methionine + 7Me-GpppA-C5 feline coronavirus
-
 S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
 ?
2.1.1.57 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
-
 ?
2.1.1.57 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
-
 ?
2.1.1.57 S-adenosyl-L-methionine + GpppA-Cn Dengue virus exclusive methylation at the 2'O position S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.56 Dengue virus
-
-
-
2.1.1.57 Dengue virus
-
-
-
2.1.1.57 feline coronavirus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.57
-
 Dengue virus
2.1.1.57 Sepharose resin column chromatography and Superdex 200 gel filtration feline coronavirus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.56 S-adenosyl-L-methionine + G(5')pppR-RNA
-
 Dengue virus S-adenosyl-L-homocysteine + m7G(5')pppR-RNA
-
 ?
2.1.1.57 additional information maximum 2'O-MTase activity requires the cap and at least 6 nucleotides Dengue virus ?
-
 ?
2.1.1.57 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 ?
-
 ?
2.1.1.57 S-adenosyl-L-methionine + 7Me-GpppA-C5
-
 feline coronavirus S-adenosyl-L-homocysteine + 7Me-GpppA-2'OMe-C5
-
 ?
2.1.1.57 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
-
 ?
2.1.1.57 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
-
 ?
2.1.1.57 S-adenosyl-L-methionine + GpppA-Cn exclusive methylation at the 2'O position Dengue virus S-adenosyl-L-homocysteine + GpppA-2'OMe-Cn
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
2.1.1.56 mRNA cap (guanine N-7) methyltransferase part of the NS5 protein Dengue virus
2.1.1.56 mRNA cap (guanine N-7) MTase part of the NS5 protein Dengue virus
2.1.1.56 nonstructural protein 5 methyltransferase
-
 Dengue virus
2.1.1.56 NS5MTase the protein contains mRNA cap (guanine N-7) methyltransferase and (nucleoside-2'O)-methyltransferase activities Dengue virus
2.1.1.57 (nucleoside-2'O)-methyltransferase part of the NS5 protein Dengue virus
2.1.1.57 2'O-MTase
-
 Dengue virus
2.1.1.57 2'O-MTase
-
 feline coronavirus
2.1.1.57 nonstructural protein 16
-
 feline coronavirus
2.1.1.57 nonstructural protein 16 methyltransferase
-
 feline coronavirus
2.1.1.57 nonstructural protein 5 methyltransferase
-
 Dengue virus
2.1.1.57 NS5MTase the protein contains mRNA cap (guanine N-7) methyltransferase and (nucleoside-2'O)-methyltransferase activities Dengue virus
2.1.1.57 nsp16
-
 feline coronavirus
2.1.1.57 RNA (nucleoside-2'O)-methyltransferase
-
 feline coronavirus
2.1.1.57 S-adenosyl-L-methionine-dependent mRNA cap MTase
-
 feline coronavirus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.57 7.5
-
-
 feline coronavirus
2.1.1.57 8
-
-
 Dengue virus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.57 7 9
-
 Dengue virus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.1.1.57 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
2.1.1.57 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

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.1.1.57 0.00034
-
 in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus S-adenosyl-L-homocysteine
2.1.1.57 0.00063
-
 in 40 mM Tris-HCl, pH 7.5, at 30°C Dengue virus sinefungin
2.1.1.57 0.144
-
-
 feline coronavirus S-adenosyl-L-homocysteine