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Literature summary for 3.4.21.91 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
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Cloned(Commentary)

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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

Organism UniProt Comment Textmining
West Nile virus
-
-
-
West Nile virus NY99
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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

Synonyms Comment Organism
NS2B-NS3 proteinase
-
 West Nile virus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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]

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

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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
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