色谱

色谱

• • 上一篇    下一篇

双配体磁性氢键有机框架材料的制备及用于糖基化肽与磷酸化肽的同时富集

杨艺涵1, 成澄宇1, 刘彦均1, 肖兆晴1, 仰俊伟1, 李松芮1,2, 郑海娇1,2*, 贾琼1,2*   

  1. 1.吉林大学化学学院，长春 130012;
    2.吉林大学国家级化学实验教学示范中心，长春 13001
  • 收稿日期:2026-01-29
  • 通讯作者: * 郑海娇.Tel: (0431)85095144, E-mail: hjzheng@jlu.edu.cn. 博士, 副教授, 主要从事蛋白组学分析研究.* 贾琼.Tel: (0431)85095144, E-mail: jiaqiong@jlu.edu.cn. 博士, 教授, 主要从事复杂样品分离分析研究.
  • 基金资助:
    国家重大科研仪器开发专项(22327804)，吉林省教育厅科学技术研究项目(JJKH20241244KJ).

Dual-ligand hydrogen-bonded organic frameworks: Tailored for glycopeptides and phosphopeptides analysis

YANG Yihan1, Cheng Chengyu1, Liu Yanjun1, XIAO Zhaoqing1, YANG Junwei1, LI Songrui1,2, ZHENG Haijiao1,2*, JIA Qiong1,2*   

  1. 1. College of Chemistry, Jilin University, Changchun 130012, China;
    2. National Demonstration Center for Experimental Chemistry Education, Jilin University, Changchun 130012, China
  • Received:2026-01-29
  • Supported by:
    National Major Research Instrument Development Program (22327804), and Science and Technology Research Project of Education Department of Jilin Province (JJKH20241244KJ).

PDF

39

摘要： 蛋白质糖基化和磷酸化均是生命体中重要的翻译后修饰，它们不仅影响着蛋白质的空间构象、生物活性、运输和定位，而且在细胞生长和分裂、信号转导、代谢途径等特定生物过程中发挥着至关重要的作用。本研究制备了一种基于尿素和苯硼酸衍生物构建的双配体氢键有机框架（hydrogen-bonded organic frameworks，HOFs）材料，并用于复杂实际样品中糖基化肽（糖肽）与磷酸化肽（磷肽）的同时富集。所合成的双配体HOFs利用尿素配体中的N-H键与1,3,5-三（4-苯硼酸）苯配体中硼酸根的氧原子形成两组N-H···O氢键，构成蜂窝状二维网状结构的HOFs，更重要的是，硼原子具有空的p轨道，能与尿素中的N形成B←N配位键，增强了HOFs框架结构的稳定性。根据理论计算，HOFs的功能位点显示出与糖肽和磷肽的协同匹配效应。另外，建立了基于该双配体磁性HOFs的磁性固相萃取-质谱分析（MSPE-MS）平台，以标准蛋白酶解物为模型，实现了该方法对糖肽和磷肽的同时富集（检出限分别低至0.4和0.1 fmol/μL）。最后，该方法在人唾液和血清样本中对于低丰度糖肽和磷肽分析中展现出较好的选择性及分离能力，为高通量糖基化及磷酸化蛋白质组学分析提供新的思路。

关键词: 氢键有机框架材料, 磁性固相萃取, 糖基化, 磷酸化, 质谱分析

Abstract: Protein glycosylation and phosphorylation stand as pivotal post-translational modifications (PTMs) in living organisms, profoundly influencing protein spatial conformation, biological activity, transportation, and localization. Moreover, they play a crucial role in specific biological processes such as cell growth and division, signal transduction, and metabolic pathways. In this study, we developed a dual-ligand hydrogen-bonded organic frameworks (HOFs) based on urea and phenylboronate salts for the simultaneous enrichment of glycopeptides and phosphopeptides from complex samples. The synthesized dual-ligand HOFs utilizes the N-H bond from the urea ligand and the oxygen atom from borate group of the 1,3,5-tris(4-phenylboronic acid)benzene ligand to form two sets of N-H···O hydrogen bonds, constructing a honeycomb-like two-dimensional network structure of HOFs. More importantly, the boron atom, with its vacant p-orbital, can form a B←N coordinate bond with the nitrogen in urea, thereby enhancing the structural stability of the HOFs. According to theoretical calculations, the functional sites of the HOFs exhibit a synergistic matching effect with both glycopeptides and phosphopeptides. We established a magnetic solid-phase microextraction-mass spectrometry (MSPE-MS) platform based on this dual-ligand magnetic HOFs. Using standard protein digests as a model, we investigated the dual enrichment capability of this method for glycopeptides and phosphopeptides, with limits of detection as low as 0.4 and 0.1 fmol/μL, respectively. Finally, this method exhibited effective separation performance for low-abundance phosphopeptides and glycopeptides in human saliva and serum samples, offering promising prospects for providing novel insights into high-throughput glycosylation and phosphorylation proteomics analysis.

Key words: hydrogen-bonded organic frameworks, dual-ligand, glycopeptides, phosphopeptides, magnetic solid-phase microextraction-mass spectrometry