Patent ID: 6333167
Filing Date: 2001-12-25
Classification: C07K,C12N,C12Q,G01N

Abstract:
A method of evaluating a compound for the ability to inhibit proteolysis of a proteolytic substrate, comprising:(a) culturing a recombinant eukaryotic host cell which expresses (i) a chimeric protein comprising a transcription repressor portion and a protease recognition portion, said protease recognition portion comprising a proteolytic cleavage site of a proteolytic substrate of interest, wherein the chimeric polypeptide is attached to a non-nuclear membrane, and (ii) a reporter gene operably linked to a transcription regulatory sequence which is responsive to said transcription repressor;(b) adding a test compound to the recombinant host cell of step (a); and(c) monitoring expression of a reporter gene product, wherein an increase in expression in the presence of the test compound added in step (b) over expression in the absence of the test compound is an indication that the test compound inhibits proteolysis of the proteolytic substrate.