<br>双功能 Ti(IV)-IMAC 材料可同时富集和分离多种糖肽和磷酸肽,Analytical Chemistry

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双功能 Ti(IV)-IMAC 材料可同时富集和分离多种糖肽和磷酸肽
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-06-08 , DOI: 10.1021/acs.analchem.1c01324
Junfeng Huang ₁ , Xiaoyan Liu ₂ , Danqing Wang ₃ , Yusi Cui ₃ , Xudong Shi ₄ , Jing Dong ₂ , Mingliang Ye ₂ , Lingjun Li _{1,

3}

Affiliation

对同一生物样品中具有不同翻译后修饰 (PTM) 的多种蛋白质/肽进行同时富集和分级分离，对于减少样品消耗、避免复杂的样品处理并能够研究不同 PTM 之间的潜在串扰是非常必要的。在这项工作中，我们报告了一种使用双功能 Ti(IV)-IMAC 材料同时富集和分离糖肽、磷酸肽和 6-磷酸甘露糖 (M6P) 糖肽的新方法。此外，我们还根据中性糖肽和唾液酸糖肽以及单磷酸肽和多磷酸肽的静电或亲水特性的差异进行不同的洗脱过程，从而使它们的分离成为可能。这些分离有效且高效地消除了用于唾液酸糖肽检测的中性糖肽的信号抑制，从而允许将单磷酸肽与多磷酸肽分离，以及检测不含上述修饰的M6P糖肽。与传统方法相比，该新策略将糖肽、磷酸肽和M6P糖肽的覆盖率和识别数分别提高了1.9、2.3和4.3倍。这是第一篇关于通过双功能 Ti(IV)-IMAC 同时富集和分离中性和唾液酸糖肽、单磷酸肽和多磷酸肽以及 M6P 糖肽的报告，揭示了这些重要 PTM 之间潜在串扰的新见解。

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Dual-Functional Ti(IV)-IMAC Material Enables Simultaneous Enrichment and Separation of Diverse Glycopeptides and Phosphopeptides

Simultaneous enrichment and fractionation of diverse proteins/peptides possessing different post-translational modifications (PTMs) from the same biological samples is highly desirable to reduce sample consumption, avoid complicated sample processing, and enable studies of potential crosstalks between different PTMs. In this work, we report a new approach to enable simultaneous enrichment and separation of glycopeptides, phosphopeptides, and mannose-6-phosphate (M6P) glycopeptides by using a dual-functional Ti(IV)-IMAC material. Moreover, we also made the separation of neutral and sialyl glycopeptides and mono- and multi-phosphopeptides possible by performing different elution processes according to the differences in their electrostatic or hydrophilic properties. These separations are effective and efficient to eliminate the signal suppression from neutral glycopeptides for sialyl glycopeptide detection, allowing separation of mono-phosphopeptides from multi-phosphopeptides, as well as detection of M6P glycopeptides that are free from the abovementioned modifications. This new strategy significantly improves the coverage and identification numbers of glycopeptides, phosphopeptides, and M6P glycopeptides by 1.9, 2.3, and 4.3-fold compared with the conventional method, respectively. This is the first report on simultaneous enrichment and separation of neutral and sialyl glycopeptides, mono- and multi-phosphopeptides, and M6P glycopeptides via dual-functional Ti(IV)- IMAC, revealing novel insights into potential crosstalk among these important PTMs.

更新日期：2021-06-22

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