Capture of Phosphopeptides Using α-Zirconium Phosphate Nanoplatelets

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• 作者：Songyun Xu ; John C. Whitin ; Tom To-Sang Yu ; Houjiang Zhou ; Dazhi Sun ; Hung-Jue Sue ; Hanfa Zou ; Harvey J. Cohen ; Richard N. Zare
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：July 15, 2008
• 年：2008
• 卷：80
• 期：14
• 页码：5542 - 5549
• 全文大小：213K
• 年卷期：v.80,no.14(July 15, 2008)
• ISSN：1520-6882

文摘

α-Zirconium phosphate nanoplatelets (α-ZrPN) were studied as a binding agent for phosphopeptides. Nanoplatelets of α-zirconium phosphate were incubated overnight with zirconium oxychloride, followed by centrifugation, and washed twice with water followed by an aqueous solution of 80% acetonitrile to form the binding agent. α-ZrPN were able specifically to capture phosphoserine-containing peptides from a tryptic digest of a complex peptide mixture in which its abundance was only 0.05%. α-ZrPN also bound peptides containing phosphothreonine and phosphotyrosine. The limit of detection for phosphopeptides is ~2 fmol, based on using matrix-assisted laser desorption/ionization mass spectrometry. α-ZrPN were applied for the analysis of tryptic digests of mouse liver and leukemia cell phosphoproteomes and succeeded in identifying 158 phosphopeptides (209 phosphorylation sites) from 101 phosphoproteins in mouse liver lysate and 78 phosphopeptides (104 phosphorylation sites) from 59 phosphoproteins in leukemia cell extract. For these two tryptic digests, the α-ZrPN approach is able to capture more phosphopeptides than that obtained from TiO₂ particles or from Fe³⁺-IMAC beads, but each method is able to bind some phosphopeptides that the others do not.