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Literature summary extracted from

  • Kamkaew, M.; Chimnaronk, S.
    Characterization of soluble RNA-dependent RNA polymerase from dengue virus serotype 2: The polyhistidine tag compromises the polymerase activity (2015), Protein Expr. Purif., 112, 43-49.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.7.48 pharmacology the viral RNA polymerase is an attractive target for inhibition in the treatment of viral infections Dengue virus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.48 bacterial production of the recombinant DENV-2 RdRp was difficult due to its low expression and solubility levels, recombinant expression of three different forms of DENV-2 RdRp as either N- or C-terminally His-tagged fusions, or without tag in Escherichia coli strains Rosetta (DE3) and BL21 (DE3). The presence of both the N- and C-terminal His-tag has a deleterious effect on polymerase activity, overview Dengue virus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.48 additional information
-
 additional information steady-state kinetics analysis of non-tagged DENV-2 RdRp in the presence of constant poly(rC) and Mg2+ concentrations Dengue virus
2.7.7.48 2.39
-
 GTP pH 7.5, 30°C, recombinant nontagged enzyme Dengue virus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.7.48 soluble
-
 Dengue virus
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.48 Mg2+ required Dengue virus
2.7.7.48 additional information recombinant non-tagged RdRp does not require manganese ions to activate RNA polymerization Dengue virus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.48 60000
-
-
 Dengue virus
2.7.7.48 73200
-
 recombinant nontagged enzyme, gel filtration Dengue virus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.48 nucleoside triphosphate + RNAn Dengue virus
-
 diphosphate + RNAn+1
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.48 Dengue virus P29991 polyprotein; serotype 2, a flavivirus
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.48 three different recombinant forms of DENV-2 RdRp as either N- or C-terminally His-tagged fusions, or without tag from Escherichia coli strain s Rosetta (DE3) and BL21 (DE3) to homogeneity by nickel affinity and anion exchange chromatography, followed by gel filtration and ultracentrifugation Dengue virus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.48 GTP + RNAn with poly(rC) Dengue virus diphosphate + RNAn+1
-
 ?
2.7.7.48 nucleoside triphosphate + RNAn
-
 Dengue virus diphosphate + RNAn+1
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.7.48 monomer 1 * 60000, recombinant nontagged enzyme, SDS-PAGE Dengue virus

Synonyms

EC Number Synonyms Comment Organism
2.7.7.48 RDRP
-
 Dengue virus
2.7.7.48 RNA-dependent RNA polymerase
-
 Dengue virus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.48 30
-
 assay at Dengue virus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.48 0.345
-
 GTP pH 7.5, 30°C, recombinant nontagged enzyme Dengue virus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.48 7.5
-
 assay at Dengue virus

General Information

EC Number General Information Comment Organism
2.7.7.48 physiological function the RNA-dependent RNA polymerase activity resides in the C-terminal two-thirds of non-structural protein (NS) 5 responsible for the de novo synthesis of the viral RNA genome. RdRp specifically binds to the viral UTR Dengue virus