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Literature summary for 3.4.21.91 extracted from

  • Shiryaev, S.A.; Kozlov, I.A.; Ratnikov, B.I.; Smith, J.W.; Lebl, M.; Strongin, A.Y.
    Cleavage preference distinguishes the two-component NS2B-NS3 serine proteinases of Dengue and West Nile viruses (2007), Biochem. J., 401, 743-752.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.21.91

Application

Application Comment Organism
drug development unique cleavage preferences of the Dengue virus protease promise novelty for the design of potent inhibitors that may be selective for the flaviviral proteases over proteases of the host cell and thus provide a solid base for follow-on structural and functional studies Dengue virus type 2
drug development unique cleavage preferences of the West Nile virus protease promise novelty for the design of potent inhibitors that may be selective for the flaviviral proteases over proteases of the host cell and thus provide a solid base for follow-on structural and functional studies West Nile virus

Cloned(Commentary)

Cloned (Comment) Organism
into the pET101 and expressed in Escherichia coli BL21(DE3) Codon Plus cells West Nile virus

Protein Variants

Protein Variants Comment Organism
H51A devoid of any proteolytic activity and incapable of autolytically cleaving either the NS2B sequence or the NS3pro-hel boundary. Resistant to the proteolysis in trans by the external, highly active NS2B-NS3pro construct West Nile virus
K48A mutation of the C-terminal amino acid residue of the NS2B sequence, inactivates the autolytic cleavage site. Does not have any significant effect on the catalytic activity. Readily cleaved in cis by the integral NS2B-NS3pro activity West Nile virus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.036
-
 t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin mutant K48A West Nile virus
0.04
-
 t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin wild-type West Nile virus
0.059
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin mutant K48A West Nile virus
0.071
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin wild-type West Nile virus

Organism

Organism UniProt Comment Textmining
Dengue virus type 2
-
-
-
Dengue virus type 2 16681
-
-
-
West Nile virus
-
-
-
West Nile virus NY99
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant DV NS2B-NS3pro, C-terminally tagged with a hexahistidine tag, purified by Co2+ affinity chromatography Dengue virus type 2
wild-type and mutant NS2B-NS3pro and NS2B-NS3prohel constructs, C-terminally tagged with a hexahistidine tag, purified by Co2+ affinity chromatography West Nile virus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information strict substrate specificity, the (K/R)(K/R)R-GG amino acid motif is optimal. Efficiently cleaves peptides derived from the West Nile virus and Dengue virus capsid protein C, from the NS2A/NS2B junction region and NS4B/NS5 junction region and from NS2B/NS3 West Nile virus ?
-
 ?
additional information strict substrate specificity, the (K/R)(K/R)R-GG amino acid motif is optimal. Efficiently cleaves peptides derived from the West Nile virus and Dengue virus capsid protein C, from the NS2A/NS2B junction region and NS4B/NS5 junction region and from NS2B/NS3 West Nile virus NY99 ?
-
 ?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus ?
-
 ?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
 Dengue virus type 2 ?
-
 ?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
 Dengue virus type 2 16681 ?
-
 ?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus NY99 ?
-
 ?
t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus ?
-
 ?
t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus NY99 ?
-
 ?

Synonyms

Synonyms Comment Organism
NS2B-NS3 protease
-
 West Nile virus
NS2B-NS3 protease
-
 Dengue virus type 2
NS2B-NS3 serine proteinase
-
 West Nile virus
NS2B-NS3 serine proteinase
-
 Dengue virus type 2

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.18
-
 t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin mutant K48A West Nile virus
0.22
-
 t-butoxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin wild-type West Nile virus
5.3
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin mutant K48A West Nile virus
6.3
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin wild-type West Nile virus