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Literature summary 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

Activating Compound

EC Number Activating Compound Comment Organism Structure
5.6.2.4 ATP catalytic DNA helicase activity coupled with NTPase stimulated by Hepacivirus C
5.6.2.4 polyadenylate doubling of ATPase activity in the presence of Hepacivirus C
5.6.2.4 polyuridylate doubling of ATPase activity in the presence of, lowers Km for the ATP substrate Hepacivirus C

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.4.13 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
5.6.2.4 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

EC Number Inhibitors Comment Organism Structure
3.6.4.13 2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 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
3.6.4.13 2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-O-ATP and N1-OH-ITP Hepacivirus C
3.6.4.13 GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 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
3.6.4.13 KCl slight decrease of activity in presence of Hepacivirus C
3.6.4.13 N1-O-ATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 N1-OH-ITP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 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
5.6.2.4 2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-oxoadenosine 5'-triphosphate and N1-hydroxyinosine 5'-triphosphate Hepacivirus C
5.6.2.4 GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 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
5.6.2.4 KCl slight decrease of activity in presence of Hepacivirus C
5.6.2.4 N1-hydroxyinosine 5'-triphosphate inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 N1-O-ATP
-
 Hepacivirus C
5.6.2.4 N1-OH-ITP
-
 Hepacivirus C
5.6.2.4 N1-oxoadenosine 5'-triphosphate inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6.4.13 0.163
-
 ATP addition of polyuridylate lowers Km for the ATP substrate Hepacivirus C
3.6.4.13 0.256
-
 ATP wild-type Hepacivirus C
5.6.2.4 0.163
-
 ATP addition of polyuridylate lowers Km for the ATP substrate Hepacivirus C
5.6.2.4 0.256
-
 ATP wild-type Hepacivirus C

Metals/Ions

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.6.4.13 ATP + H2O Hepacivirus C NTPase activity analyzed, ambiguous helicase activity, enzyme capable for unwinding RNA and DNA ADP + phosphate
-
 ?
5.6.2.4 ATP + H2O Hepacivirus C catalytic DNA helicase activity is coupled with NTPase and is stimulated by ATP ADP + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.6.4.13 Hepacivirus C
-
-
-
5.6.2.4 Hepacivirus C
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.4.13 gel filtration, recombinant protein Hepacivirus C
5.6.2.4 gel filtration, recombinant protein Hepacivirus C

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.6.4.13 additional information
-
 structure of nucleic base and ribose fragment of NTP molecule has a slight effect on inhibitory properties Hepacivirus C
5.6.2.4 additional information
-
 DNA helicase reaction can proceed in two modes depending on the ratio between enzyme and substrate concentration, non-catalytic in the case of enzyme excess and catalytic in the case of tenfold substrate excess, structure of nucleic base and ribose fragment of NTP molecule has a slight effect on inhibitory properties, duplex DNA oligonucleotides used for determination of DNA helicase activity Hepacivirus C

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.6.4.13 ATP + H2O NTPase activity analyzed, ambiguous helicase activity, enzyme capable for unwinding RNA and DNA Hepacivirus C ADP + phosphate
-
 ?
5.6.2.4 ATP + H2O catalytic DNA helicase activity is coupled with NTPase and is stimulated by ATP Hepacivirus C ADP + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.6.4.13 non structural protein 3 ambiguous Hepacivirus C
3.6.4.13 protein NS3 ambiguous Hepacivirus C
5.6.2.4 non structural protein 3 ambiguous Hepacivirus C
5.6.2.4 protein NS3 ambiguous Hepacivirus C

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.6.2.4 37
-
 activity assay at Hepacivirus C

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.6.2.4 7.4
-
 activity assay at Hepacivirus C

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.6.4.13 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
3.6.4.13 0.109
-
 N1-OH-ITP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.116
-
 2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.116
-
 2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.141
-
 3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.145
-
 beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-O-ATP and N1-OH-ITP Hepacivirus C
3.6.4.13 0.205
-
 N1-O-ATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.26
-
 3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.277
-
 2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.291
-
 2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.298
-
 2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.443
-
 3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.576
-
 GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 0.721
-
 2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 1.3
-
 ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 1.46
-
 UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
3.6.4.13 5
-
 AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 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
5.6.2.4 0.109
-
 N1-OH-ITP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.116
-
 2'-dTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.116
-
 2',3'-ddATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.141
-
 3'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.145
-
 beta,gamma-methylene-ATP efficient inhibitor, like the N1-oxides N1-oxoadenosine 5'-triphosphate and N1-hydroxyinosine 5'-triphosphate Hepacivirus C
5.6.2.4 0.205
-
 N1-O-ATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.26
-
 3'-dUTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.277
-
 2'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.291
-
 2'-dATP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.298
-
 2',3'-ddTTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.443
-
 3'-dGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.576
-
 GTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 0.721
-
 2',3'-ddGTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 1.3
-
 ADP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 1.46
-
 UTP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C
5.6.2.4 5
-
 AMP inhibition of NTPase activity of NS3 protein by NTP derivatives Hepacivirus C