Torta Federico, Fusi Matteo, Casari Carlo S, Bassi Andrea Li, Bachi Angela
Division of Genetics & Cell Biology, San Raffaele Scientific Institute, Milan, Italy.
Methods Mol Biol. 2011;790:173-81. doi: 10.1007/978-1-61779-319-6_13.
Alterations in protein phosphorylation, a posttranslational modification (PTM) that regulates many -processes in living cells, is a fundamental mechanism of many diseases, including cancer. Phosphoproteomics, with the combined use of affinity chromatography and electrospray ionization (ESI) or matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, is shedding light into phosphorylation signaling pathways at the proteome level and helps to solve difficulties related to sample complexity and phosphopeptide enrichment. One of the most frequent and efficient methods used to enrich samples for the phosphorylated components is titanium dioxide chromatography. Titanium dioxide has a high affinity for phosphopeptides and can also be selective in specific experimental conditions. Here, we describe a protocol for the use of a MALDI plate covered with titanium dioxide nanostructured film, a device developed for a rapid and efficient study of phosphorylated peptides.
蛋白质磷酸化是一种调节活细胞中许多过程的翻译后修饰（PTM），其改变是包括癌症在内的许多疾病的基本机制。磷酸化蛋白质组学结合使用亲和色谱和电喷雾电离（ESI）或基质辅助激光解吸/电离（MALDI）质谱，正在蛋白质组水平上揭示磷酸化信号通路，并有助于解决与样品复杂性和磷酸肽富集相关的难题。用于富集样品中磷酸化成分的最常用且有效的方法之一是二氧化钛色谱法。二氧化钛对磷酸肽具有高亲和力，并且在特定实验条件下也具有选择性。在此，我们描述了一种使用覆盖有二氧化钛纳米结构薄膜的MALDI板的方案，该装置是为快速有效地研究磷酸化肽而开发的。
