Patent ID: 6165731
Filing Date: 2000-12-26
Classification: C07K,C12Q,G01N

Abstract:
A method of identifying a compound that affects an activity of a polypeptide subunit of a Skp1p/Cullin/F-box ubiquitin ligase (SCF) complex, said method comprising:(a) contacting a test sample comprising a chimeric SCF complex assembled from subunits derived from yeast or human and another species and an ubiquitin-conjugating enzyme (CDC34p) with a test compound under suitable conditions that allow the components to interact;(b) adding to the components of (a) an ubiquitin-activating enzyme E1, ubiquitin, adenosine tri-phosphate (ATP), and a SCF substrate; and(c) comparing the ubiquitination of said SCF substrate in the test sample to ubiquitination of said SCF substrate in a sample not contacted with said test compound, wherein the difference in ubiquitination of said SCF substrate in the test sample is indicative of the effect of said compound on said polypeptide subunits.