智能聚合物基材料富集磷酸化肽和糖肽的研究进展

翻译后修饰是蛋白质组学研究的前沿和重点,它不仅调节着蛋白质的折叠、状态、活性、定位以及蛋白质间的相互作用,也能帮助科学家更全面地了解生物体的生命过程,为疾病的预测、诊断和治疗提供更加强大的支撑和依据。翻译后修饰产物(例如磷酸化肽和糖肽)丰度很低,且存在着强烈的背景干扰,很难直接用质谱进行分析,因此迫切需要开发高效的富集材料和技术来选择性富集翻译后修饰产物。近年来,智能聚合物基材料通过外部物理、化学或生物刺激可逆地改变其结构和功能,实现对磷酸化肽和糖肽高度可控的吸附和脱附,进而衍生开发出一系列新颖的富集方法,极大地吸引研究者们的兴趣。一方面,智能聚合物基材料的响应变化包括材料疏水性的增加或减少、形状和形貌的改变、表面电荷的重新分布以及亲和配体的暴露或隐藏等特性。这些特性使得目标物和智能聚合物基材料之间的亲和力可以通过简单改变外部条件(如温度、pH值、溶剂极性和生物分子等)实现更可控和更智能的精细调节。另一方面,智能聚合物基材料为集成功能模块提供了便捷的可扩展平台,例如特定的识别组件,显著提高了目标物质的分离选择性。智能聚合物基材料在分离方面展现出巨大的潜力,这为蛋白质翻译后修饰产物的分析和研究带来了希望。围绕上述主题,该文依据Web of Science近20年来近50篇代表性文献,概述了智能聚合物基材料在磷酸化肽和糖肽分离及富集中的发展方向。

Advances in enrichment of phosphorylated peptides and glycopeptides by smart polymer-based materials

Protein post-translational modification(PTM)is at the forefront of focus of proteomics research.It not only regulates protein folding,state,activity,localization,and protein interactions,but also helps scientists understand the biological processes of organisms more comprehensively,providing stronger support and basis for the prediction,diagnosis,and treatment of diseases.In living organisms,there are more than 300 types of PTMs of proteins and their modification processes are dynamic.At the same time,protein modifications do not exist in isolation.The occurrence of the same physiological or pathological process requires the joint action of various modified proteins,which affect and coordinate with each other.Owing to the low abundance of PTM products(e.g.,phosphorylated peptides or glycopeptides)and the presence of strong background interference,it is difficult to analyze them directly through mass spectrometry.Therefore,the development efficient materials and techniques for the selective enrichment of PTM peptides is urgently needed.Conventional separation methods have partially solved the challenges involved in the enrichment of glycopeptides and phosphorylated peptides;however,there are some inevitable issues,such as the excessive binding force of metal ions(e.g.,Fe3+and Ti^4+)toward multiple phosphorylated peptides,resulting in difficulty in elution and identification through mass spectrometry.In addition,owing to the insufficient binding affinity of materials toward glycopeptides,most glycopeptides that have been identified at present are of the sialic acid type,and a large number of neutral glycans,for instance,O-link glycopeptides and high mannose-type glycans are difficult to enrich and identify.The emergence of smart polymers provides a new avenue for the development of PTMenriched materials.Several studies have reported that smart polymers can reversibly change their structure and function through external physical,chemical,or biological stimulation,to achieve highly controllable adsorption and desorption of phosphorylated peptides and glycopeptides.Based on this strategy,a series of novel enrichment materials and methods have been developed,which have greatly attracted the interest of researchers.On the one hand,the response changes of smart polymers include the increase or decrease of hydrophobicity,the change of shape and morphology,the redistribution of surface charge,the exposure or hiding of affinity ligands,etc.Changes in these properties can be achieved by simply changing external conditions such as temperature,pH,solvent polarity,and biomolecules.These properties,in turn,enable the fine-tuning of the affinity between the target and the smart polymers.Furthermore,the affinity can provide an additional driving force,which can significantly improve biological separation.On the other hand,smart polymers provide a series of convenient and expandable platforms for integrating various functional modules,such as specific recognition components,which will facilitate the development of novel enrichment materials for protein methylation,acetylation,and ubiquitination.Smart polymer materials show great potential in the field of separation,which is promising for the analysis and research of protein PTMs.This review summarizes the research progress of smart polymer materials for the separation and enrichment of phosphorylated peptides and glycopeptides according to nearly 50 representative articles from the Web of Science in the past two decades.