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Literature summary for 3.4.21.91 extracted from

  • Shiryaev, S.A.; Aleshin, A.E.; Ratnikov, B.I.; Smith, J.W.; Liddington, R.C.; Strongin, A.Y.
    Expression and purification of a two-component flaviviral proteinase resistant to autocleavage at the NS2B-NS3 junction region (2007), Protein Expr. Purif., 52, 334-339.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
analysis NS2B-NS3pro K48A mutant construct is well suited for follow-up purification and structural and drug design studies West Nile virus

Cloned(Commentary)

Cloned (Comment) Organism
NS2B-NS3pro wild-type and K48A constructs cloned into pET101 vector and expressed in Escherichia coli BL21 (DE3) Codon Plus cells West Nile virus

Protein Variants

Protein Variants Comment Organism
K48A is a single-chain protein and fully resistant to autolysis at the linker region, exhibits high stability and functional activity, cleaves pyroglutamic acid-Arg-Thr-Lys-Arg-7-amino-4-methylcoumarin West Nile virus

Inhibitors

Inhibitors Comment Organism Structure
Aprotinin completely inhibits autolysis of the NS2B-NS3pro construct West Nile virus
decanoyl-Arg-Val-Lys-Arg-chloromethylketone completely inhibits autolysis of the NS2B-NS3pro construct West Nile virus
nona-D-Arg-amide completely inhibits autolysis of the NS2B-NS3pro construct West Nile virus

Organism

Organism UniProt Comment Textmining
West Nile virus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
NS2B-NS3pro wild-type and K48A constructs purified by Co2+ affinity chromatography and gel filtration, to homogeneity West Nile virus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information during purification, incomplete autolytic conversion of the 36 kDa NS2B-NS3pro construct into the individual 30 kDa NS3pro and the NS2B sequence (5-6 kDa). Undergoes self-cleavage during expression and purification at the K48G-G site of the glycine linker West Nile virus ?
-
?
pyroglutamic acid-Arg-Thr-Lys-Arg-7-amido-4-methylcoumarin + H2O
-
West Nile virus ?
-
?

Synonyms

Synonyms Comment Organism
NS2BNS3(pro)tease
-
West Nile virus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000006
-
nona-D-Arg-amide
-
West Nile virus
0.000025
-
Aprotinin
-
West Nile virus
0.0004
-
decanoyl-Arg-Val-Lys-Arg-chloromethylketone
-
West Nile virus