Ultra Acidic Strong Cation Exchange Enabling the Efficient Enrichment of Basic Phosphopeptides

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• 作者：Marco L. Hennrich ; Henk W. P. van den Toorn ; Vincent Groenewold ; Albert J. R. Heck ; Shabaz Mohammed
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：February 21, 2012
• 年：2012
• 卷：84
• 期：4
• 页码：1804-1808
• 全文大小：211K
• 年卷期：v.84,no.4(February 21, 2012)
• ISSN：1520-6882

文摘

We present a straightforward method to enrich phosphopeptides with multiple basic residues, an under-represented class in common enrichment strategies. Our method is based on a two-dimensional strong cation exchange (SCX) strategy, operating at two different acidic pHs, enabling both separation and enrichment of different classes of phosphopeptides. The principle of enrichment is based on the change of net charge of phosphorylated peptides under strong acidic conditions in the second SCX, whereas the net charge of regular peptides remains unchanged, thus enabling separation based on net charge. Application of our tandem SCX approach to a modest amount of human cells allowed the identification of over 10鈥?00 unique 鈥渂asic鈥?phosphopeptides of which many represent putative targets of basophilic kinases.