CELL-FREE NOTCH CLEAVAGE ANALYSIS METHOD AND DRUG SCREENING METHOD

摘要

The present invention provides a method for analyzing cell-free Notch cleavage which can be utilized for drug screening of ³-secretase inhibitors effective for the treatment of Alzheimer's disease and the drug screening method, is the method for analyzing cell-free Notch cleavage having a cell-free cleavage reaction step of contacting a crude membrane fraction of cells in which a Notch protein mutant has been expressed with a subject substance and a detection step of detecting a fragment of an amino terminal side and a fragment of a carboxyl terminal side produced by intramembrane proteolysis (dual sequential cleavage in putative transmembrane domain) of the Notch protein mutant in the above cell free cleavage reaction step, wherein the effect of the subject substance on the intramembrane proteolysis of the Notch protein mutant is analyzed, and the Notch protein mutant has at least the putative transmembrane domain of a Notch protein, deletes a part of an amino acid sequence at the amino terminal side than at least the putative transmembrane domain and has antibody recognition sites at the amino terminal side than the putative transmembrane domain and the carboxyl terminal side, respectively.