Xu Dongpo, Yan Guoquan, Gao Mingxia, Deng Chunhui, Zhang Xiangmin
Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China.
Anal Bioanal Chem. 2017 Feb;409(6):1607-1614. doi: 10.1007/s00216-016-0101-0. Epub 2016 Nov 29.
Glycosylation and phosphorylation as the commonest and most important posttranslational modifications of proteins play critical roles in biological processes. Specific and sensitive strategies have been developed to identify the glycosylation and phosphorylation of proteins by mass spectrometry but most of these methods have mainly focused on enriching glycopeptides or phosphopeptides separately, and few of them can enrich both of them. In this work, highly selective SiO-NH@TiO hollow microspheres were designed and synthesized for the simultaneous selective enrichment of both phosphopeptides and glycopeptides. Because of the bifunctionalized property of the titanium dioxide and the amino group, the hollow microspheres were successfully applied for the simultaneous enrichment of phosphopeptides and glycopeptides. We evaluated the enrichment selectivity of the SiO-NH@TiO hollow microspheres by capturing phosphopeptides and glycopeptides from a peptide mixture of bovine β-casein, horseradish peroxidase, and bovine serum albumin in a molar ratio of 1:1:500 (1.7 × 10 mol β-casein and horseradish peroxidase, 8.5 × 10 mol bovine serum albumin in 100 μL). Graphical Abstract Workflow of the simultaneous enrichment strategy for phosphopeptides and glycopeptides by SiO-NH@TiO hollow microspheres.
糖基化和磷酸化作为蛋白质最常见且最重要的翻译后修饰，在生物过程中发挥着关键作用。已开发出特异性和灵敏性的策略，通过质谱法鉴定蛋白质的糖基化和磷酸化，但这些方法大多主要集中于分别富集糖肽或磷酸肽，很少有方法能同时富集两者。在本研究中，设计并合成了高选择性的SiO-NH@TiO中空微球，用于同时选择性富集磷酸肽和糖肽。由于二氧化钛和氨基的双功能化特性，中空微球成功应用于同时富集磷酸肽和糖肽。我们通过从摩尔比为1:1:500（100 μL中含1.7×10⁻⁵ mol β-酪蛋白和辣根过氧化物酶、8.5×10⁻⁴ mol牛血清白蛋白）的牛β-酪蛋白、辣根过氧化物酶和牛血清白蛋白的肽混合物中捕获磷酸肽和糖肽，评估了SiO-NH@TiO中空微球的富集选择性。图摘要：SiO-NH@TiO中空微球同时富集磷酸肽和糖肽的策略工作流程。
