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Literature summary extracted from

  • Shiryaev, S.A.; Radichev, I.A.; Ratnikov, B.I.; Aleshin, A.E.; Gawlik, K.; Stec, B.; Frisch, C.; Knappik, A.; Strongin, A.Y.
    Isolation and characterization of selective and potent human Fab inhibitors directed to the active-site region of the two-component NS2B-NS3 proteinase of West Nile virus (2010), Biochem. J., 427, 369-376.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.91 expressed in Escherichia coli CodonPlus (DE3)-RIPL cells West Nile virus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.91 G22S no activity West Nile virus
3.4.21.91 H51A no activity West Nile virus
3.4.21.91 K48A the mutant displays slightly increased catalytic efficiency compared to the wild type enzyme West Nile virus
3.4.21.91 R76L the mutant displays reduced catalytic efficiency compared to the wild type enzyme West Nile virus
3.4.21.91 T52V the mutant displays increased catalytic efficiency compared to the wild type enzyme West Nile virus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.21.91 antibody D05320
-
 West Nile virus
3.4.21.91 antibody D05321
-
 West Nile virus
3.4.21.91 antibody D05322
-
 West Nile virus
3.4.21.91 antibody D05323
-
 West Nile virus
3.4.21.91 antibody D05444
-
 West Nile virus
3.4.21.91 antibody D05445
-
 West Nile virus
3.4.21.91 antibody D05446
-
 West Nile virus
3.4.21.91 Aprotinin
-
 West Nile virus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.91 0.0013
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme T52V, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.0588
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme K48A, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.071
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.089
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme R76L, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.91 West Nile virus
-
-
-
3.4.21.91 West Nile virus NY99
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.91 HiTrap Co2+-chelating column chromatography, gel filtration West Nile virus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.91 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.21.91 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus NY99 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg + 7-amino-4-methylcoumarin
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.21.91 NS2B-NS3 proteinase
-
 West Nile virus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.91 0.013
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme T52V, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 5.25
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme K48A, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 6.3
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 7.3
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme R76L, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.21.91 0.00002
-
 Aprotinin wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.000031
-
 antibody D05323 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.000035
-
 antibody D05445 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.000058
-
 antibody D05444 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.00017
-
 antibody D05322 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.000264
-
 antibody D05320 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.000288
-
 antibody D05446 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 0.0004
-
 antibody D05321 wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.21.91 82
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme R76L, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 88
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin wild type enzyme, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 89.3
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme K48A, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus
3.4.21.91 100
-
 L-pGlu-L-Arg-L-Thr-L-Lys-L-Arg-7-amido-4-methylcoumarin mutant enzyme T52V, in 20 mM Tris/HCl buffer, pH 8.0, at 22°C West Nile virus