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Literature summary for 3.4.21.91 extracted from

  • Rut, W.; Zhang, L.; Kasperkiewicz, P.; Poreba, M.; Hilgenfeld, R.; Drag, M.
    Extended substrate specificity and first potent irreversible inhibitor/activity-based probe design for Zika virus NS2B-NS3 protease (2017), Antiviral Res., 139, 88-94 .
    View publication on PubMed
Search for more literature for 3.4.21.91

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00418
-
 Ac-Arg-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
0.0042
-
 Ac-D-Arg-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
0.006
-
 Ac-D-Arg-Lys-Orn-Arg(Me)-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
0.00799
-
 Ac-D-Lys-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
0.01385
-
 Ac-Nle-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus

Organism

Organism UniProt Comment Textmining
Zika virus A0A024B7W1 ZIKV
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ac-Arg-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin + H2O
-
 Zika virus ?
-
 ?
Ac-D-Arg-Lys-Orn-Arg(Me)-7-amido-4-carbamoylmethylcoumarin + H2O
-
 Zika virus ?
-
 ?
Ac-D-Arg-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin + H2O substrate profiling of the P1 position using Ac-Ala-Arg-Leu-X-7-amido-4-carbamoylmethylcoumarin (X is natural or unnatural amino acid) Zika virus ?
-
 ?
Ac-D-Lys-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin + H2O
-
 Zika virus ?
-
 ?
Ac-Nle-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin + H2O
-
 Zika virus ?
-
 ?
additional information analysis of substrate specificity at substrate positions P1, P2, P3, and P4, and cleavage site parameters, detailed overview. Enzyme cleavage sequence is Ac-Ala-Arg-Leu-X-7-amido-4-carbamoylmethylcoumarin (X is a natural or nonnatural amino acid). The P4-P1 optimal sequence is D-Arg-Lys-Orn-Arg. Most preferred amino acid residues at P1 are L-Arg(Me) and L-Arg. At P2, the NS2B-NS3 protease displays also a very high level of specificity. The most preferred amino acids are the non-proteinogenic L-Orn (100%) and L-Arg(Z)2 (40%). Natural amino acids interact significantly weaker with the S2 pocket, and only L-Lys and L-Arg are recognized (38% and 17%, respectively). Similarly to the S2 and S1 pockets, the S3 site exhibits very narrow specificity. L-Lys is the most preferentially recognized residue at P3 among all tested amino acids. Other amino acids such as L-Orn, L-Arg, L-Arg(Z)2, L-hArg, L-Agp or L-Phe(guan) are recognized with significantly lower affinity. However, all these amino acids have basic side chains. The S4 site exhibits very broad substrate specificity. Almost all amino acids, including unnatural ones, are recognized at the same level at P4 position Zika virus ?
-
 ?

Synonyms

Synonyms Comment Organism
NS2B-NS3 protease
-
 Zika virus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Zika virus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.08
-
 Ac-Arg-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
4.4
-
 Ac-Nle-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
4.64
-
 Ac-D-Arg-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
5.43
-
 Ac-D-Arg-Lys-Orn-Arg(Me)-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
7.99
-
 Ac-D-Lys-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Zika virus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
258.37
-
 Ac-Arg-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
317.69
-
 Ac-Nle-Lys-Lys-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
905
-
 Ac-D-Arg-Lys-Orn-Arg(Me)-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
1000
-
 Ac-D-Lys-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus
1104.76
-
 Ac-D-Arg-Lys-Orn-Arg-7-amido-4-carbamoylmethylcoumarin pH 8.5, 22°C Zika virus