Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden.
Anal Chem. 2010 Jun 1;82(11):4577-83. doi: 10.1021/ac100589j.
A novel on-target phosphopeptide enrichment method is presented that allows specific enrichment and direct analysis by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) of phosphorylated peptides. Spots consisting of a thin film of anatase titanium dioxide are sintered onto a conductive glass surface. Enrichment and analysis can be performed on the modified target with minimal sample handling. The protocol leads to an enrichment efficiency that is superior to what has been reported before for similar methods. The method was tested using beta-casein as a model phosphorylated protein as well as with a custom peptide mixed with its phosphorylated form. A very low detection limit, a significantly improved phosphoprofiling capability, and a simple experimental approach provide a powerful tool for the enrichment, detection, and analysis of phosphopeptides.
提出了一种新颖的针对靶磷酸肽富集方法，该方法允许通过基质辅助激光解吸电离质谱（MALDI-MS）对磷酸化肽进行特异性富集和直接分析。将一层锐钛矿二氧化钛的薄膜烧结到导电玻璃表面上。可以对修饰后的靶进行最小化的样品处理，从而进行富集和分析。该方案导致的富集效率优于以前报道的类似方法。该方法使用β-酪蛋白作为模型磷酸化蛋白以及与混合其磷酸化形式的定制肽进行了测试。非常低的检测限，大大改善的磷酸化分析能力以及简单的实验方法为磷酸肽的富集，检测和分析提供了强大的工具。
