Substrate inhibition kinetic model for West Nile virus NS2B-NS3 protease

Suzanne M. Tomlinson, Stanley Watowich

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

West Nile virus (WNV) has recently emerged in North America as a significant disease threat to humans and animals. Unfortunately, no approved antiviral drugs exist to combat WNV or other members of the genus Flavivirus in humans. The WNV NS2B-NS3 protease has been one of the primary targets for anti-WNV drug discovery and design since it is required for virus replication. As part of our efforts to develop effective WNV inhibitors, we reexamined the reaction kinetics of the NS2B-NS3 protease and the inhibition mechanisms of newly discovered inhibitors. The WNV protease showed substrate inhibition in assays utilizing fluorophore-linked peptide substrates GRR, GKR, and DFASGKR. Moreover, a substrate inhibition reaction step was required to accurately model kinetic data generated from protease assays with a peptide inhibitor. The substrate inhibition model suggested that peptide substrates could bind to two binding sites on the protease. Reaction product analogues also showed inhibition of the protease, demonstrating product inhibition in addition to and distinct from substrate inhibition. We propose that small peptide substrates and inhibitors may interact with protease residues that form either the P3-P1 binding surface (i.e., the S3-S1 sites) or the P1′-P3′ interaction surface (i.e., the S1′-S3′ sites). Optimization of substrate analogue inhibitors that target these two independent sites may lead to novel anti-WNV drugs.

Original languageEnglish (US)
Pages (from-to)11763-11770
Number of pages8
JournalBiochemistry
Volume47
Issue number45
DOIs
StatePublished - Nov 11 2008

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West Nile virus
Viruses
Peptide Hydrolases
Kinetics
Substrates
Peptides
Assays
Flavivirus
Drug Design
Drug Discovery
Virus Replication
North America
Fluorophores
Antiviral Agents
Reaction products
Reaction kinetics
Binding Sites
Animals
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry

Cite this

Substrate inhibition kinetic model for West Nile virus NS2B-NS3 protease. / Tomlinson, Suzanne M.; Watowich, Stanley.

In: Biochemistry, Vol. 47, No. 45, 11.11.2008, p. 11763-11770.

Research output: Contribution to journalArticle

Tomlinson, Suzanne M. ; Watowich, Stanley. / Substrate inhibition kinetic model for West Nile virus NS2B-NS3 protease. In: Biochemistry. 2008 ; Vol. 47, No. 45. pp. 11763-11770.
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