Abstract:
PURPOSE: A method for identifying protein kinase substrate specificity using a peptide library is provided to rapidly determining the substrate specificity of a protein phosphatase, and to examine the role of the protein phosphatase. CONSTITUTION: A method for identifying protein kinase substrate specificity comprises the following steps: forming a peptide library with a constant amino acid sequence on the surface of a solid support resin; forming a hybrid with randomized peptide libraries by repeating the previous step; reacting the peptide libraries with a phosphatase for forming the peptide library with phosphorylated threonine or serine; reacting an antibody combined with a biotin with the phosphorylated peptide library; reacting the obtained product with neutravidin or streptavidin including phosphatase or peroxidase; detecting the change of properties by the reaction of the phosphatase or the peroxidase for selecting the solid support resin with transformed peptide libraries; and an analyzing the amino acid sequence of the peptide after separating the peptide from the resin.