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Literature summary for 3.4.21.91 extracted from

  • Radichev, I.; Shiryaev, S.A.; Aleshin, A.E.; Ratnikov, B.I.; Smith, J.W.; Liddington, R.C.; Strongin, A.Y.
    Structure-based mutagenesis identifies important novel determinants of the NS2B cofactor of the West Nile virus two-component NS2B-NS3 proteinase (2008), J. Gen. Virol., 89, 636-641.
    View publication on PubMed
Search for more literature for 3.4.21.91

Application

Application Comment Organism
drug development useful for the development of specific allosteric inhibitors designed to interfere with the productive interactions of NS2B with NS3pro West Nile virus

Cloned(Commentary)

Cloned (Comment) Organism
NS2B-NS3pro construct cloned into the pET101/D-TOPO vector C-terminally tagged with the His x 6 tag and expressed in Escherichia coli BL21(DE3) Codon Plus cells West Nile virus

Protein Variants

Protein Variants Comment Organism
D42G has no dramatic effect on either the catalytic activity (50% of wild-type) or self-proteolysis of NS2B-NS3pro. Aprotinin efficiently inhibits proteolytic activity West Nile virus
G22S resistant to autoproteolysis at the flexible linker region, retains 3% residual activity West Nile virus
K48A resistant to autoproteolysis at the flexible linker region. Aprotinin efficiently inhibits proteolytic activity West Nile virus
additional information D(32)DD(34) substitution for AAA inactivates NS2B-NS3pro, retains 1% residual activity West Nile virus

Inhibitors

Inhibitors Comment Organism Structure
Aprotinin
-
 West Nile virus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.004
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin mutant G22S West Nile virus
0.004
-
 butyloxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin mutant G22S West Nile virus
0.03
-
 pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin wild-type West Nile virus
0.03
-
 butyloxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin wild-type West Nile virus

Organism

Organism UniProt Comment Textmining
West Nile virus P06935
-
-
West Nile virus NY99 P06935
-
-

Purification (Commentary)

Purification (Comment) Organism
NS2B-NS3pro construct purified by Co2+-chelating Sepharose Fast Flow chromatography West Nile virus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
butyloxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus butyloxycarbonyl-Arg-Val-Arg-Arg + 7-amino-4-methylcoumarin
-
 ?
butyloxycarbonyl-Arg-Val-Arg-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus NY99 butyloxycarbonyl-Arg-Val-Arg-Arg + 7-amino-4-methylcoumarin
-
 ?
myelin basic protein + H2O
-
 West Nile virus ?
-
 ?
myelin basic protein + H2O
-
 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-amino-4-methylcoumarin
-
 ?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
 West Nile virus NY99 pyroglutamic acid-Arg-Thr-Lys-Arg + 7-amino-4-methylcoumarin
-
 ?

Synonyms

Synonyms Comment Organism
NS2B-NS3 proteinase
-
 West Nile virus

Cofactor

Cofactor Comment Organism Structure
NS2B protein the G(22) residue and the negatively charged patch D(32)DD(34) of NS2B are part of an important configuration required for NS2B-NS3pro activity West Nile virus