期刊
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
卷 15, 期 16, 页码 1481-1488出版社
WILEY
DOI: 10.1002/rcm.394
关键词
-
A new method combining chemical modification and affinity purification is described for the characterization of serine and threonine phosphopeptides in proteins. The method is based on the conversion of phosphoserine and phosphothreonine residues to S-(2-mercaptoethyl)cysteinyl or beta -methyl-S-(2-mercaptoethyl)cysteinyl residues by beta -elimination/1,2-ethanedithiol addition, followed by reversible biotinylation of the modified proteins. After trypsin digestion, the biotinylated peptides were affinity-isolated and enriched, and subsequently subjected to structural characterization by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Database searching allowed for automated identification of modified residues that were originally phosphorylated. The applicability of the method is demonstrated by the identification of all known phosphorylation sites in a mixture of alpha -casein, beta -casein, and ovalbumin. The technique has potential for adaptations to proteome-wide analysis of protein phosphorylation. Copyright (C) 2001 John Wiley & Sons, Ltd.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据