Optimized Protocol for On-Target Phosphopeptide Enrichment Prior to Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry Using Mesoporous Titanium Dioxide
Optimized Protocol for On-Target Phosphopeptide Enrichment Prior to Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry Using Mesoporous Titanium Dioxide
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DOI：
10.1021/ac100589j
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发表时间：
2010-06-01
影响因子：
7.4
通讯作者：
Hernandez, Victor Agmo
中科院分区：
文献类型：
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作者：
Eriksson, Anna;Bergquist, Jonas;Hernandez, Victor Agmo
提出了一种新颖的靶向磷酸肽富集方法，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。由锐钛矿二氧化钛薄膜组成的斑点被烧结到导电玻璃表面上。可以用最少的样品处理对修改后的目标进行富集和分析。该协议的富集效率优于之前报道的类似方法。使用 β-酪蛋白作为模型磷酸化蛋白以及与其磷酸化形式混合的定制肽对该方法进行了测试。极低的检测限、显着提高的磷酸化分析能力和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。
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 phosphotylated 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.