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Literature summary for 3.6.4.13 extracted from
- Analysis of the nucleoside triphosphatase, RNA triphosphatase, and unwinding activities of the helicase domain of dengue virus NS3 protein (2009), FEBS Lett., 583, 691-696.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Poly(rU) | stimulates the ATPase activity of NS3 | Dengue virus |  |
| polyU | stimulatory effect of polyU on ATP hydrolysis is significantly attenuated when NaCl concentration is 50 mM or higher, functional binding of polyU mainly through electrostatic interaction, binding triggers a conformational rearrangement that activates the catalytic core of the enzyme for ATP hydrolysis | Dengue virus | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli, recombinant protein | Dengue virus |
| expression of NS3 ATPase/helicase in Escherichia coli, expression of the His-tag C-terminal domain | Dengue virus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K199A/T200A | site-directed mutagenesis, mutant avoid of basal and of RNA-stimulated NTPase activity | Dengue virus |
| K199A/T200A | site-directed mutagenesis, the mutation in the C-terminal domain of NS3 eliminates both the basal and the RNA-stimulated NTPase activity | Dengue virus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| KCl | - |
Dengue virus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | activates, preferred metal ion | Dengue virus | |
| Mg2+ | activity depends on divalent cations, assay concentration 1.5 mM, rate of ATP hydrolysis 10 times enhanced with Mg2+ as divalent cation cofactor, rate of ATP hydrolysis increases slightly when the NaCl concentration is elevated in the range of 10 mM and 200 mM | Dengue virus | |
| Mn2+ | activates | Dengue virus | |
| Mn2+ | activity depends on divalent cations, assay concentration 1.5 mM, rate of ATP hydrolysis more than 100 times over basal levels with Mn2+ as divalent cation cofactor | Dengue virus | |
| additional information | the ATPase activity requires a divalent cation cofactor to function but is not sensitive to high ionic strength | Dengue virus | |
| NaCl | slightly activating ATPase activity at 10-200 mM, and RTPase at 15-50 mM | Dengue virus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | Dengue virus | - |
ADP + phosphate | - |
? | |
| ATP + H2O | Dengue virus PL046 | - |
ADP + phosphate | - |
? | |
| RNA + H2O | Dengue virus | - |
? | - |
? | |
| RNA + H2O | Dengue virus PL046 | - |
? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Dengue virus | - |
- |
- |
| Dengue virus | - |
serotype 2, PL046 strain | - |
| Dengue virus PL046 | - |
serotype 2, PL046 strain | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tag C-terminal domain of NS3 protein from Escherichia coli by nickel affinity chromatography | Dengue virus |
| recombinant protein, gel filtration, SDS-PAGE | Dengue virus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
biochemical properties and enzymatic activity of the RNA-helicase domain, functional characterization to get information about the flavivirus replication mechanism, NTPase-deficient mutant generated, RNA binding features, electrostatic interaction with RNA, basal ATPase activity insensitive to high ionic strength | Dengue virus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | - |
Dengue virus | ADP + phosphate | - |
? | |
| ATP + H2O | NS3 C-terminal domain catalyzes ATP hydrolysis in the presence of MgCl2 or MnCl2. MgCl2 is more effective than MnCl2 at inducing ATPase activity at concentrations ranging from 0.1 mM to 5 mM. ATP hydrolysis is required for the unwinding activity of DENV NS3H | Dengue virus | ADP + phosphate | - |
? | |
| ATP + H2O | wild-type and mutant, NTPase activity analyzed, functional binding of RNA analyzed | Dengue virus | ADP + phosphate | - |
? | |
| ATP + H2O | - |
Dengue virus PL046 | ADP + phosphate | - |
? | |
| ATP + H2O | NS3 C-terminal domain catalyzes ATP hydrolysis in the presence of MgCl2 or MnCl2. MgCl2 is more effective than MnCl2 at inducing ATPase activity at concentrations ranging from 0.1 mM to 5 mM. ATP hydrolysis is required for the unwinding activity of DENV NS3H | Dengue virus PL046 | ADP + phosphate | - |
? | |
| ATP + H2O | wild-type and mutant, NTPase activity analyzed, functional binding of RNA analyzed | Dengue virus PL046 | ADP + phosphate | - |
? | |
| additional information | the helicase domain of Dengue virus NS3 protein, i.e. DENV NS3H, contains RNA-stimulated nucleoside triphosphatase, NTPase, ATPase/helicase, and RNA 5'-triphosphatase, RTPase, activities that are essential for viral RNA replication and capping. A 5'-tailed RNA is a better RTPase substrate than an RNA containing no 5'-dangling nucleotide | Dengue virus | ? | - |
? | |
| additional information | the helicase domain of Dengue virus NS3 protein, i.e. DENV NS3H, contains RNA-stimulated nucleoside triphosphatase, NTPase, ATPase/helicase, and RNA 5'-triphosphatase, RTPase, activities that are essential for viral RNA replication and capping. A 5'-tailed RNA is a better RTPase substrate than an RNA containing no 5'-dangling nucleotide | Dengue virus PL046 | ? | - |
? | |
| RNA + H2O | - |
Dengue virus | ? | - |
? | |
| RNA + H2O | helicase/unwinding activity, ATP hydrolysis is required for the unwinding activity of DENV NS3H | Dengue virus | ? | - |
? | |
| RNA + H2O | - |
Dengue virus PL046 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | the C-terminal domain of the enzyme contains the Walker A and Walker B motifs, i.e. motif I, GK(S/T) and motif II, DExD/H | Dengue virus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ATPase/helicase | - |
Dengue virus |
| DENV NS3H | - |
Dengue virus |
| helicase | - |
Dengue virus |
| NS3 ATPase/helicase | - |
Dengue virus |
| NS3 protein | - |
Dengue virus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Dengue virus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Dengue virus |
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