| 摘要 |
The present invention relates to a process for the identification of a kinase substrate specificity by using a peptide library. More particulary, the present invention is directed to a process for indent ifying a kinase substrate specificity, comprising: i ) linking a plurality of peptides, having a same amino acid sequence, to the surface of a solid support resin to obtain a peptide library which has peptides, with the same amino acid sequence, on the surface of the solid support resin; ii) repeating the step i ) with changing the amino acid sequence to give a random mixture of peptide libraries, wherein each solid suppot resin has peptides with a same amino acid sequence, however the amino acid sequence liked to each solid support is different from one another," iii) phosphorylating selectively a specific amino acid of a peptide which has a certain amino acid sequence by reacting the random mixture of the peptide libraries with a kinase, to give a phosphorylated peptide library; iv) reacting the phosphorylated peptide library with an antibody which is conjugated with one of a phosphatase and a peroxidase and binds selectively to a phosphorylated amino acid, followed by the addition of a substrate selectively acting on the thus prepared reaction mixture; v) selecting a solid support resin which has a peptide library of which property has been changed, by detecting the change of property due to the reaction of the sbustrate with one of the phosphatase and the peroxidase; and vi) separating a peptide from the selected solid support resin followed by analysing a sequece of the separated peptide. |
