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Literature summary for 3.6.4.13 extracted from

  • Mukovnya, A.V.; Tunitskaya, V.L.; Khandazhinskaya, A.L.; Golubeva, N.A.; Zakirova, N.F.; Ivanov, A.V.; Kukhanova, M.K.; Kochetkov, S.N.
    Hepatitis C virus helicase/NTPase: an efficient expression system and new inhibitors (2008), Biochemistry (Moscow), 73, 660-668.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli, strains XL-1 Blue, Rosetta (DE3), M15 (pREP4), vector pET-21-2c, kinetics of NS3 protein accumulation upon its expression in Escherichia coli at 25°C for 1-5 h shown Hepacivirus C

Inhibitors

Inhibitors Comment Organism Structure
2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
2'-deoxythymidine 5'-phosphoryl-beta,gamma-hypophosphate i.e. ppopT, dTTP analogue, most efficient inhibitor of NTPase activity among nucleotide derivaties, inhibits the ATP-dependent helicase reaction and also the ATP-independent duplex unwinding, structure of nucleic base and ribose fragment of NTP molecule have a slight effects on inhibitory properties Hepacivirus C
2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-O-ATP and N1-OH-ITP Hepacivirus C
GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
Imidodiphosphate maximal inhibitory activity among diphosphate analogues, non-catalytic and catalytic conditions, inhibits the ATP-dependent helicase reaction but no effect on the ATP-independent duplex unwinding, structure of nucleic base and ribose fragment of NTP molecule have a slight effects on inhibitory properties Hepacivirus C
KCl slight decrease of activity in presence of Hepacivirus C
N1-O-ATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
N1-OH-ITP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.163
-
ATP addition of polyuridylate lowers Km for the ATP substrate Hepacivirus C
0.256
-
ATP wild-type Hepacivirus C

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activity 3-5-fold lower when magnesium ions are replaced by Hepacivirus C
Mg2+ maximal NTPase activity achieved in the presence of 1.5-2 mM MgCl2 Hepacivirus C
Mn2+ activity 3-5-fold lower when magnesium ions are replaced by Hepacivirus C
Ni2+ activity 3-5-fold lower when magnesium ions are replaced by Hepacivirus C
Zn2+ activity 3-5-fold lower when magnesium ions are replaced by Hepacivirus C

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + H2O Hepacivirus C NTPase activity analyzed, ambiguous helicase activity, enzyme capable for unwinding RNA and DNA ADP + phosphate
-
?

Organism

Organism UniProt Comment Textmining
Hepacivirus C
-
-
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration, recombinant protein Hepacivirus C

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
structure of nucleic base and ribose fragment of NTP molecule has a slight effect on inhibitory properties Hepacivirus C

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O NTPase activity analyzed, ambiguous helicase activity, enzyme capable for unwinding RNA and DNA Hepacivirus C ADP + phosphate
-
?

Synonyms

Synonyms Comment Organism
non structural protein 3 ambiguous Hepacivirus C
protein NS3 ambiguous Hepacivirus C

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.097
-
2'-deoxythymidine 5'-phosphoryl-beta,gamma-hypophosphate i.e. ppopT, dTTP analogue, inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.109
-
N1-OH-ITP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.116
-
2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.116
-
2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.141
-
3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.145
-
beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-O-ATP and N1-OH-ITP Hepacivirus C
0.205
-
N1-O-ATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.26
-
3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.277
-
2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.291
-
2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.298
-
2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.443
-
3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.576
-
GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
0.721
-
2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
1.3
-
ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
1.46
-
UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5
-
AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C