Profiling of Endogenous Serum Phosphorylated Peptides by Titanium (IV) Immobilized Mesoporous Silica Particles Enrichment and MALDI-TOFMS Detection
Profiling of Endogenous Serum Phosphorylated Peptides by Titanium (IV) Immobilized Mesoporous Silica Particles Enrichment and MALDI-TOFMS Detection
复制标题
通过钛 (IV) 固定介孔二氧化硅颗粒富集和 MALDI-TOFMS 检测分析内源血清磷酸化肽
DOI：
10.1021/ac801974f
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发表时间：
2009-01-01
影响因子：
7.4
通讯作者：
Zou, Hanfa
中科院分区：
文献类型：
--
作者：
Hu, Lianghai;Zhou, Houjiang;Zou, Hanfa
磷酸化是蛋白质最重要的翻译后修饰之一，它调节蛋白质的多种生物学功能和活性。蛋白质的磷酸化也与癌细胞的通路有关。我们之前利用高度有序的介孔二氧化硅颗粒，基于尺寸排阻和吸附机制的结合，从人血浆中富集了低分子量蛋白质组。在此，用膦酸钛修饰高度有序的介孔二氧化硅颗粒，以选择性地从复杂的肽和蛋白质混合物中捕获磷酸肽。根据 MALDI-TOFMS 检测，人血清中添加的 β-酪蛋白磷酸肽和标准磷酸肽的检测限低至 1.25 fmol。修饰的介孔二氧化硅颗粒进一步用于富集肝细胞癌患者和健康个体血清中的磷酸肽，然后用 MALDI-TOFMS 进行分析。进一步应用同量异位标记与 MALDI-TOFMS/MS 相对和绝对定量标记相结合来验证 MALDI-TOFMS 的血清磷酸肽分析结果。癌症患者和健康人之间的血清磷酸肽谱存在显着差异，这表明该技术在癌症诊断和生物标志物发现方面的潜在能力。这里开发的方法适用于其他生物样本和多种疾病。
Phosphorylation is one of the most important post-translational modifications of proteins, which modulates a wide range of biological functions and activities of proteins. The phosphorylation of proteins is also associated with the pathway of cancer cells. We have previously enriched the low molecular weight proteome from human plasma based on the combination of size exclusion and adsorption mechanism by using highly ordered mesoporous silica particles. Herein, highly ordered mesoporous silica particles were modified with titanium phosphonate to selectively capture the phosphopeptides from complex peptide and protein mixtures. The limit of detection for phosphopeptides from beta-casein and standard phosphopeptide spiked in human serum was as low as 1.25 fmol based on MALDI-TOFMS detection. The modified mesoporous silica particles were further used to enrich phosphopeptides from serum of hepatocellular carcinoma patients and healthy individuals and then analyzed with MALDI-TOFMS. The combination of isobaric tagging for relative and absolute quantitation labeling with MALDI-TOFMS/MS was further applied to validate the serum phosphopeptide profiling result of MALDI-TOFMS. The profiling of the serum phosphopeptides between the cancer patients and healthy persons was distinguishingly different, which indicated the potential ability of this technique for cancer diagnosis and biomarker discovery. The approach developed here would be applicable to other biological samples and a wide variety of diseases.