Literature summary for 2.7.7.48 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.

Application

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

Cloned(Commentary)

Cloned (Comment)	Organism
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

Cloned (Comment)

Organism

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]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
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
2.39	-	GTP	pH 7.5, 30°C, recombinant nontagged enzyme	Dengue virus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
soluble	-	Dengue virus	-	-

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

Purification (Comment)	Organism
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)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
RDRP	-	Dengue virus
RNA-dependent RNA polymerase	-	Dengue virus

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

General Information

General Information	Comment	Organism
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