发明名称 |
Protein kinase peptide substrate determination using peptide libraries |
摘要 |
<p>A method of isolating and identifying peptide substrates for a protein kinase is disclosed. The method involves a combination of size exclusion and gallium-based metal affinity chromatography. The method includes the steps of incubating a protein kinase with a peptide library in the presence of kinase reaction components, the library comprising library members; separating library members from the kinase reaction components using size exclusion chromatography to give a pool of phosphopeptides and unphosphorylated peptides; contacting the pool with immobilized gallium ions to form chelated phosphopeptides; eluting chelated phosphopeptides away from the gallium ions to give eluted phosphopeptides; sequencing the eluted phosphopeptides, whereby a preferred amino acid sequence of a preferred peptide substrate for a protein kinase is elucidated. Also disclosed is a method of identifying a compound that modulates the protein kinase catalyzed phosphorylation of a peptide substrate and a method of designing protein kinase substrates. <IMAGE></p> |
申请公布号 |
EP1361285(A2) |
申请公布日期 |
2003.11.12 |
申请号 |
EP20020077062 |
申请日期 |
2002.05.28 |
申请人 |
SMITHKLINE BEECHAM CORPORATION |
发明人 |
BLACKBURN, ROBERT KEVIN;BRAMSON, HAROLD NEAL;MOYER, MARY BENBOW;STUART, JAMES DARREN |
分类号 |
G01N33/48;C07K7/08;C12N9/00;C12Q1/48;C40B40/02;C40B50/06;G01N30/06;G01N30/88;G01N33/15;G01N33/50;G01N33/68;(IPC1-7):C12Q1/48 |
主分类号 |
G01N33/48 |
代理机构 |
|
代理人 |
|
主权项 |
|
地址 |
|