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Literature summary for 3.4.21.91 extracted from
- Switching the substrate specificity of the two-component NS2B-NS3 flavivirus proteinase by structure-based mutagenesis (2007), J. Virol., 81, 4501-4509.
Application
| Application | Comment | Organism |
|---|---|---|
| drug development | switched cleavage preferences of West Nile virus NS2B-NS3(pro)teinase may provide valuable data for the design of optimized substrates and substrate-based selective inhibitors of flaviviral proteinases | West Nile virus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| wild-type Dengue virus and West Nile virus NS2B-NS3pro, together with West Nile virus/Dengue virus chimeras expressed with C-terminal His tags in Escherichia coli BL21(DE3) Codon Plus cells | Dengue virus |
| wild-type Dengue virus and West Nile virus NS2B-NS3pro, together with West Nile virus/Dengue virus chimeras expressed with C-terminal His tags in Escherichia coli BL21(DE3) Codon Plus cells | West Nile virus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| P131K/T132P | decrease in kcat, which is partially compensated by an improvement in the substrate binding. Shift of the cleavage preferences toward that of the Dengue virus proteinase | West Nile virus |
| R76L | kinetic parameters are similar to those of the wild-type. Shift of the cleavage preferences toward that of the Dengue virus proteinase | West Nile virus |
| T52V | exhibits a significant loss of kcat, which is compensated by an improvement in substrate binding. Does not cause any significant differences in the cleavage subsite specificity. Cleavage preferences of the T52V mutant construct are similar to those of the original West Nile virus proteinase | West Nile virus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Aprotinin | - |
West Nile virus |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Dengue virus | - |
- |
- |
| West Nile virus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| NS2B-NS3pro constructs purified by Co2+ affinity chromatography | Dengue virus |
| NS2B-NS3pro constructs purified by Co2+ affinity chromatography | West Nile virus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| AQRRGRIG + H2O | three individual batches of the peptide results in AQRR2GRIG 3%, 2%, and 0% cleavages | West Nile virus | AQRR + GRIG | - |
? | |
| AQRRGRIG + H2O | three individual batches of the peptide results in 45%, 37%, and 36% cleavages | Dengue virus | ? | - |
? | |
| GLKRFGAK + H2O | - |
Dengue virus | GLKR + FGAK | - |
? | |
| GLKRGSAK + H2O | cleaved with 82% efficiency | Dengue virus | GLKR + GSAK | - |
? | |
| GLKRGSAK + H2O | cleaved with 9% efficiency | West Nile virus | GLKR + GSAK | - |
? | |
| GLKRTGAK + H2O | cleaved with 2% efficiency | West Nile virus | GLKR + TGAK | - |
? | |
| GLKRTGAK + H2O | cleaved with 77% efficiency | Dengue virus | GLKR + TGAK | - |
? | |
| KKQRAGVL + H2O | cleaved with 11% efficiency | Dengue virus | ? | - |
? | |
| LYKRYGGF + H2O | cleaved with 48% efficiency | Dengue virus | ? | - |
? | |
| MLKRGMPR + H2O | cleaved with 49% efficiency | Dengue virus | ? | - |
? | |
| additional information | peptides with a similar sequence, including GIKRGETD, LQKRGIVE, LSKRQHPG, LVTAGHGQ, and MLKKGMTT, are completely resistant to proteolysis | Dengue virus | ? | - |
? | |
| additional information | peptides with a similar sequence, including GIKRGETD, LQKRGIVE, LSKRQHPG, LVTAGHGQ, and MLKKGMTT, are completely resistant to proteolysis. Peptides GLKRFGAK, KKQRAGVL, LYKRYGGF, MLKRGMPR, and NQKRYGGF are also not cleaved | West Nile virus | ? | - |
? | |
| NQKRYGGF + H2O | cleaved with 7% efficiency | Dengue virus | ? | - |
? | |
| NRKR + GGPA + H2O | two individual batches of the peptide results in 78 and 77% cleavages | West Nile virus | NRKRGGPA | - |
? | |
| pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O | efficiently cleaved | West Nile virus | ? | - |
? | |
| QRRRGGTA + H2O | two individual batches of the peptide results in 58% cleavage | West Nile virus | QRRR + GGTA | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NS2B-NS3(pro)teinase | - |
Dengue virus |
| NS2B-NS3(pro)teinase | - |
West Nile virus |
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