Pieroni, L., Santolini, E., Fipaldini, C., Pacini, L., Migliaccio, G., La Monica, N.
Notes: This article describes an in vitro assay to activate hepatitis C virus (HCV) NS2-3 protease posttranslationally and methods studying the effects of several common inhibitors on enzymatic activity. HCV N2-3 protease was expressed using the Rabbit Reticulocyte Lysate System in the presence of [35S]methionine, and the NS2-3 protease was activated by detergent cleavage. The protein was incubated with various proteinase inhibitors (aprotinin, antipain, DTT, E64, pepstatin, EGTA, EDTA, iodoacetamide, N-ethylaleimide, 1,10-phenanthroline, phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, TLCK and TPCK). (0533)