发明名称 |
Protein kinase peptide substrate determination using peptide libraries |
摘要 |
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.
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申请公布号 |
US2003113711(A1) |
申请公布日期 |
2003.06.19 |
申请号 |
US20020155481 |
申请日期 |
2002.05.24 |
申请人 |
BLACKBURN ROBERT KEVIN;BRAMSON HAROLD NEAL;MOYER MARY BENBOW;STUART JAMES DARREN |
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):C12Q1/70;G01N33/53 |
主分类号 |
G01N33/48 |
代理机构 |
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主权项 |
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地址 |
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