Mass spectrometry analysis of phosphopeptides after peptide carboxy group derivatization.
Mass spectrometry analysis of phosphopeptides after peptide carboxy group derivatization.
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DOI：
10.1021/ac801220c
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发表时间：
2008-09
影响因子：
7.4
通讯作者：
Yawei Xu;Lijuan Zhang;Haojie Lu;Pengyuan Yang
中科院分区：
文献类型：
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作者：
Yawei Xu;Lijuan Zhang;Haojie Lu;Pengyuan Yang
提出了一种收率接近100%的肽羧基衍生化方法，大大提高了磷酸肽的离子化效率。该方法采用1-（2-嘧啶基）哌嗪（PP）为衍生试剂，具有重现性好、操作简便、反应时间快、无副反应等优点。PP衍生化提高了肽特别是磷酸肽的疏水性、p I值和气相碱性。在基质辅助激光解吸电离（MALDI）源中，四种合成磷酸肽的电离效率提高了50-101倍，三种非磷酸肽的电离效率提高了10-40倍。在电喷雾电离（ESI）源，PP衍生的磷酸肽也得到了更高的电离效率的改善比nonphosphopeptides。当这种方法被应用到更复杂的混合物，胰蛋白酶BSA底物掺入一个单一的磷酸肽在不同的摩尔比，该磷酸肽的信号强度总是有最大的增量在所有这些肽。显然，这种易于操作以及高度特异性的方法提供了一个有前途的工具，高通量磷酸化蛋白质组研究。
A nearly 100% yield peptide carboxy group derivatization method was offered to largely enhance phosphopeptide ionization efficiency. This method, adopting 1-(2-pyrimidyl) piperazine (PP) as the derivatization reagent, shows several advantages such as good reproducibility, ease of handling, rapid reaction time, and no side reaction. PP derivatization improves the hydrophobicities, p I values, and gas-phase basicities of peptides especially those of phosphopeptides. In the matrix assisted laser desorption ionization (MALDI) source, the ionization efficiencies of four synthetic phosphopeptides were increased by 50-101 times while that of three nonphosphopeptides were 10-40-fold. In the electrospray ionization (ESI) source, PP-derivatized phosphopeptides also gave much higher ionization efficiency improvements than nonphosphopeptides. When this method was applied to much more complex mixtures, tryptic BSA digests spiked with one single phosphopeptide in different molar ratios, the signal intensity of this phosphopeptide always had the largest increment among all those peptides. Obviously, this easily manipulated as well as highly specific method provides a promising tool for high-throughput phosphoproteome research.