| 摘要 |
PROBLEM TO BE SOLVED: To provide a method for isolating a peptide substrate of protein kinase, and to provide a method for identifying the substrate. SOLUTION: This method includes a combination of size exclusion chromatography and metal affinity chromatography based on gallium. A preferable peptide substrate of the protein kinase and its amino acid sequence are elucidated by incubating the protein kinase with a peptide library in the presence of a kinase reacting component, separating a library member from the kinase reacting component by size exclusion chromatography to obtain a pool of phosphopeptide and nonphosphorylated peptide, contacting the pool with a gallium ion to form a chelated phosphopeptide, eluting the chelated phosphopeptide from gallium ion and determining the sequence of the eluted phosphopeptide. The method for identifying a compound for modulating phosphorylation of the peptide substrate catalyzed by the protein kinase and a design method for the protein kinase substrate are also provided.
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