method for reproducing in vitro the proteolytic activity of hcv protease ns3 virus.

摘要

it is a methodology in order to reproduce in vitro the activities associated with the ns3 serino - protease protein of hcv.that includes the use of both sequences contained in ns3 that sequences contained in ns4a.this methodology exploits the ability of the protein of hcv ns4a or sequences contained in it.to act as a cofactor of the activities proteasica serino - or, more generally, enzyme activities associated with ns3.the optimum activity of serino - protease is obtained when ns4a is present in a molar ratio of at least 1:1 with ns3.ns3 and ns4a may also be incorporated in the reaction mixture as precursor ns3 - ns4a, as this precursor will generate, through an event autoproteolitico.quantity equimolari of ns3 and ns4a. it is also possible to change the site of cut between ns3 and ns4a in a precursor, so that ns4a remains covalentemente tied to ns3.this precursor is not proteolizzabile can subsequently be removed the sequences that do not affect the proteolytic activity of ns3.the invention also relates to a composition of matter comprising sequences contained in ns3 and ns4a.and the use of these compositions for the preparation of a wise enzymatic capable of selecting, for therapeutic purposes, comprising inhibitors of enzymatic activity associated with ns3.the figure represents plasmidici carriers used in the methodology for the activation of the protease ns3 hcv in cell culture and in vitro.