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Literature summary for 2.7.7.48 extracted from

  • Ranjith-Kumar, C.T.; Sarisky, R.T.; Gutshall, L.; Thomson, M.; Kao, C.C.
    De novo initiation pocket mutations have multiple effects on hepatitis C virus RNA-dependent RNA polymerase activities (2004), J. Virol., 78, 12207-12217.
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
D225A mutant enzyme is capable of robust de novo initiation in the presence of Mn2+, produces reduced amounts of product of 38 nt and longer that arose from template switch Hepacivirus C
R158A mutation decreases de novo initiation of RNA synthesis Hepacivirus C
R158A/R367A/R386A mutation eliminates de novo initiation product even in presence of Mn2+, but enzyme is capable of primer extension Hepacivirus C
R158A/R386A mutation eliminates de novo initiation product even in presence of Mn2+, but enzyme is capable of primer extension Hepacivirus C
R158A/R394A mutation eliminates de novo initiation product even in presence of Mn2+, but enzyme is capable of primer extension Hepacivirus C
R386A mutation decreases de novo initiation of RNA synthesis Hepacivirus C
R394A mutation decreases de novo initiation of RNA synthesis Hepacivirus C
R48A mutation decreases de novo initiation of RNA synthesis Hepacivirus C

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0035
-
GTP pH 8.2, 25°C, enzyme form DELTA21 that lacks the C-terminal 21 hydrophobic residues Hepacivirus C
5.05
-
GTP pH 8.2, 25°C, mutant enzyme Hepacivirus C
10.1
-
GTP pH 8.2, 25°C, mutant enzyme Hepacivirus C
11.25
-
GTP pH 8.2, 25°C, mutant enzyme Hepacivirus C
15.3
-
GTP pH 8.2, 25°C, mutant enzyme R158A Hepacivirus C
25.9
-
GTP pH 8.2, 25°C, mutant enzyme Hepacivirus C
33.8
-
GTP pH 8.2, 25°C, mutant enzyme Hepacivirus C

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ likely helps to stabilize the interaction between the active site and the NTPi Hepacivirus C

Organism

Organism UniProt Comment Textmining
Hepacivirus C
-
expression in Escherichia coli
-

Purification (Commentary)

Purification (Comment) Organism
enzyme expressed in Escherichia coli Hepacivirus C

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GTP + RNAn
-
Hepacivirus C diphosphate + RNAn+1
-
?
nucleoside triphosphate + RNAn
-
Hepacivirus C diphosphate + RNAn+1
-
?

Synonyms

Synonyms Comment Organism
NS5B
-
Hepacivirus C
RNA-dependent RNA polymerase
-
Hepacivirus C