The Serine Protease, Polynucleotide-stimulated - NTPase and The RNA Helicase multiple enzymatic functions of nonstructural Protein NS3 - Model Biochemical Targets essential for Replication And Maturation of Dengue and Japanese Encephalitis Virus
a) Biochemical characterization of the NS2B/NS3pro serine protease complex: structure-function relations; generation and assay of active site mutants; analysis of NS2B - NS3pro interaction by site-directed mutagenesis and chimeric constructs; proteochemometric models; inhibitor design and evaluation.
b) Biochemical characterization of the RNA helicase function of the NS3 protein; establishment of assays for NTPase/helicase activity; structure-function relations by active site mutagenesis studies; development of high-throughput assays and - if possible - design and synthesis of inhibitors.
c) Molecular cloning, expression and purification of the NS2B/NS3 proteins from Japanese encephalitis virus; biochemical characterization with protease / helicase substrates and assay development; mechanistic analysis of substrate binding and inhibitor design.
