College of Chemistry, Jilin University, Changchun 130012, China.
Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, College of Life Sciences, Jilin University, Changchun 130012, China.
J Mater Chem B. 2023 Jun 7;11(22):4874-4881. doi: 10.1039/d2tb01708c.
Protein glycosylation and phosphorylation are the two most important post-translational modifications, which play a vital role in physiological and pathological processes. Before the comprehensive characterization of glycoproteome/phosphoproteome through the mass spectrometry (MS) technique, it is necessary to perform a highly specific enrichment procedure because glycoproteins/phosphoproteins inherently occur in low abundances. Herein, we have reported a novel magnetic β-cyclodextrin-based host-guest Ti-phenolic network material, focusing on the simultaneous enrichment of glycopeptides/phosphopeptides hydrophilic interaction chromatography and immobilized metal ion chromatography. Ti ions and glutathione-derived adamantine were introduced by metal-phenolic interactions and host-guest interactions. The material possesses biocompatibility, good hydrophilicity, strong magnetic response, metal chelation effect, and demonstrates an excellent enrichment ability towards glycopeptides/phosphopeptides. Combined with MS detection, high sensitivity (0.035/0.01 fmol for IgG/β-casein) and good reusability (6 times) were achieved. In addition, its outstanding specificity was validated in quantities as low as 500 : 1 : 1 for BSA : IgG : β-casein (m/m/m). Benefiting from these merits, the adsorbent material was successfully used for the simultaneous enrichment of phosphopeptides/glycopeptides from human serum and HeLa cell lysate and can be expected to exhibit great applicability for precious and small amounts of biosamples in the study of glycoproteomics/phosphoproteomics.
蛋白质糖基化和磷酸化是两种最重要的翻译后修饰，在生理和病理过程中起着至关重要的作用。在通过质谱 (MS) 技术全面表征糖蛋白组/磷酸蛋白组之前，有必要进行高度特异性的富集程序，因为糖蛋白/磷酸蛋白的含量本来就很低。在此，我们报道了一种新型的基于磁性 β-环糊精的主体-客体 Ti-酚醛网络材料，重点是同时对糖肽/磷酸肽进行亲水相互作用色谱和固载金属离子色谱的富集。Ti 离子和来源于瓜氨酸的金刚烷通过金属-酚醛相互作用和主客体相互作用被引入。该材料具有生物相容性、良好的亲水性、强磁响应、金属螯合效应，并对糖肽/磷酸肽具有优异的富集能力。结合 MS 检测，实现了高灵敏度（0.035/0.01 fmol 用于 IgG/β-酪蛋白）和良好的可重复使用性（6 次）。此外，在 BSA/IgG/β-酪蛋白（m/m/m）低至 500：1：1 的情况下，验证了其出色的特异性。得益于这些优点，该吸附剂材料成功地用于从人血清和 HeLa 细胞裂解物中同时富集磷酸肽/糖肽，有望在糖蛋白质组学/磷酸蛋白质组学研究中对珍贵和少量生物样品表现出很好的适用性。
