METHODOLOGY TO PRODUCE, PURIFY AND ASSAY POLYPEPTIDES WITH THE PROTEOLYTIC ACTIVITY OF THE HCV NS3 PROTEASE

摘要

The process according to the present invention allows expression and isolati on of polypeptides with the proteolytic activity of HCV NS3 protease in a pure, catalytically active form, and in amounts that are sufficient for discovery of NS3 protease inhibitors and for determination of the three-dimensional structure of the NS3 protease. A further subject of the present invention is a procedure that defines the chemical and physical conditions necessary for completion of the proteolytic activity of the above polypeptides. The invention further comprises new compositions of matter (expression vectors) containing nucleotide sequences capable of expressing the above mentioned polypeptides in culture cells. Finally, new compounds of matter are defined, suitable to measure the above proteolytic activity, and useful to develop NS 3 protease inhibitors and therefore therapeutic agents for use against HCV. Th e figure shows the kinetic parameters of HCV NS3 protease using the S3 depsipeptide substrate (SEQ ID NO:45).