Quantitation of phosphopeptides using affinity chromatography and stable isotope labeling

被引：35

作者：

He, T ^{[1
]}

Alving, K ^{[1
]}

Feild, B ^{[1
]}

Norton, J ^{[1
]}

Joseloff, EG ^{[1
]}

Patterson, SD ^{[1
]}

Domon, B ^{[1
]}

机构：

[1] Celera Genom Grp, Rockville, MD 20850 USA

来源：

JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY | 2004年 / 15卷 / 03期

关键词：

D O I：

10.1016/j.jasms.2003.11.004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Reversible phosphorylation of proteins represents an important component of cellular signaling pathways. The isolation of phosphoproteins in complex mixtures and the determination of the level of phosphorylation have been and remain a major challenge. It has prompted the development of several strategies, including immobilized metal affinity capture to enrich for phosphorylated peptides. An improved methodology was published (Ficarro, et al., Nature Biotechnology 2002, 20, 301-305) that showed increased selectivity through esterification of amino acid side chain carboxylic groups of enzymatically digested peptides. This method was applied for relative quantitation of phosphopeptides in conjunction with the use of stable isotope labeling. The merits and limits of the approach are discussed and its application to the analysis of the effects of serum starvation on in vitro cultured human lung cells is presented. (C) 2004 American Society for Mass Spectrometry.

引用

页码：363 / 373

页数：11

共 44 条

[1] Selective analysis of phosphopeptides within a protein mixture by chemical modification, reversible biotinylation and mass spectrometry [J].