Patent ID: 7427478
Filing Date: 2008-09-23
Classification: A01K,A61K,C12N,H01Q

Abstract:
1. A method for identifying a compound that decreases the activity of a β-secretase polypeptide comprising steps of: (a) expressing the β-secretase polypeptide by growing a host cell transformed or transfected with a polynucleotide that encodes said polypeptide and a polynucleotide encoding a substrate of β-secretase, under conditions wherein the cell expresses the polypeptide encoded by the polynucleotide and the cell expresses the substrate of β-secretase, (b) contacting the host cell with a test compound; (c) measuring the β-secretase activity of the polypeptide in the presence and absence of the test compound,  wherein the β-secretase polypeptide comprises an amino acid sequence at least 95% identical to a fragment of the β-secretase amino acid sequence of SEQ ID NO: 43,  wherein said fragment is a continuous fragment that includes active aspartic acid catalytic sites and exhibits β-secretase activity, which is an ability to cleave a 695 isotype of APP between amino acids 596 and 597, and (d) comparing the β-secretase activity of the polypeptide expressed by the cell in the presence and absence of the test compound, wherein the β-secretase activity is determined using an antibody specific for the C-terminus of the N-terminal fragment or the N-terminus of the C-terminal fragment generated by the βsecretase cleavage of the substrate, wherein decreased β-secretase activity in the presence of the test compound identifies the test compound as a compound that decreases the β-secretase activity of the β-secretase polypeptide.