Patent ID: 6043023
Filing Date: 2000-03-28
Classification: C12Q

Abstract:
A method of identifying a substrate comprising the steps of:(a) obtaining an extract of a non-human model, wherein said extract contains at least one substrate that binds to carboxypeptidase E and said non-human model has an inactive version of carboxypeptidase E resulting in accumulation of said substrate in said non-human model;(b) fractionating said extract on an affinity resin column, wherein the affinity resin of the column comprises a protein bound to a solid support, and wherein said protein is selected from the group consisting of anhydrotrypsin, inactive carboxypeptidase E having a point mutation in the amino acid sequence of carboxypeptidase E of Glu.sup.342 to Gln.sup.342, and carboxypeptidase E used in the presence of a thiol-directed inhibitor(c) obtaining an elute fraction containing said substrate from said affinity column; and(d) purifying and identifying said substrate contained in said elute fraction.