Cloned (Comment) | Organism |
---|---|
separate recombinant expression of N-terminally His-tagged DENV virus 2 NS2B and NS3pro in Escherichia coli in inclusion bodies | Dengue virus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant linked NS2B-NS3pro, X-ray diffraction structure determination and analysis at 1.6 A resolution | Dengue virus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | separate expression of dengue virus NS3 protease and its NS2B cofactor domain and efficient co-refolding to form a stable complex, method evaluation, overview. This straightforward and robust method allows for separate isotope labeling of the two proteins, facilitating analysis by NMR spectroscopy, detailed overview. Unlinked NS2B-NS3pro behaves better in NMR spectroscopy than linked NS2B-NS3pro, which has resulted in the backbone resonance assignment of the unlinked NS2B-NS3 complex bound to a peptidic boronic acid inhibitor | Dengue virus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,6-difluoro-benzoyl-Nle-Lys-Arg-Arg-CF3-ketone | - |
Dengue virus | |
benzoyl-Nle-Lys-Arg-Arg-B(OH)2 | dynamics of NS2B and NS3pro in the presence of the peptidic inhibitor | Dengue virus | |
additional information | unlinked NS2B-NS3 complex bound to a peptidic boronic acid inhibitor | Dengue virus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dengue virus | - |
DENV2 | - |
Purification (Comment) | Organism |
---|---|
recombinant separately expressed His-tagged NS2B and NS3pro from Escherichia coli, refolded and forming the active complex, by nickel affinity chromatography, tag cleavage by thrombin and/or TEV protease, and another step of nickel affinity chromatography, and gel filtration | Dengue virus |
Renatured (Comment) | Organism |
---|---|
recombinant separately expressed His-tagged NS2B and NS3pro from Escherichia coli are co-refolded by one-step dialysis overnight at 4°C in a 2:1 M NS2B:NS3pro ratio to maximize formation of the active complex. The refolding buffer is 25 mM Tris, pH 8.5 (set at 4°C), 5% glycerol, and 100 mM NaCl | Dengue virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
benzoyl-Nle-Lys-Arg-Arg-7-amido-4-methylcoumarin + H2O | - |
Dengue virus | benzoyl-Nle-Lys-Arg-Arg + 7-amino-4-methylcoumarin | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Dengue NS3 protease | - |
Dengue virus |
NS2B-NS3 complex | - |
Dengue virus |
NS2B-NS3pro | - |
Dengue virus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Dengue virus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NS2B cofactor | essential | Dengue virus |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000019 | - |
pH 7.4, temperature not specified in the publication, versus the linked NS2B-NS3 complex | Dengue virus | 2,6-difluoro-benzoyl-Nle-Lys-Arg-Arg-CF3-ketone | |
0.000038 | - |
pH 7.4, temperature not specified in the publication, versus the linked NS2B-NS3 complex | Dengue virus | benzoyl-Nle-Lys-Arg-Arg-B(OH)2 |