蛋白质N-磷酸化修饰富集方法进展  

Progress in enrichment methods for protein N-phosphorylation

作　　者：江波[1] 高博[2] 魏淑娴 梁振[1] 张丽华[1] 张玉奎[1] JIANG Bo;GAO Bo;WEI Shuxian;LIANG Zhen;ZHANG Lihua;ZHANG Yukui(State Key Laboratory of Medical Proteomics,National Chromatographic R.&A.Center,CAS Key Laboratory of Separation Science for Analytical Chemistry,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,China;School of Chemistry and Chemical Engineering,China University of Petroleum(East China),Qingdao 266580,China)

机构地区：[1]中国科学院大连化学物理研究所,医学蛋白质组全国重点实验室,国家色谱研究中心,中国科学院分离分析化学重点实验室,辽宁大连116023 [2]中国石油大学(华东)化学化工学院,山东青岛266580

出　　处：《色谱》2024年第7期623-631,共9页Chinese Journal of Chromatography

基　　金：国家重点研发计划(2022YFC3401204);国家自然科学基金(22074140,22393931);中国科学院大连化物所创新基金(DICP I202316).

摘　　要：蛋白质磷酸化作为一种最普遍和最重要的翻译后修饰调控着几乎所有的生命过程。随着高效富集方法和生物质谱技术的快速发展,低丰度的蛋白质O-磷酸化修饰获得了规模化鉴定,从而使其生物学功能得到较为透彻的研究。而发生在组氨酸、赖氨酸和精氨酸侧链氨基的N-磷酸化修饰,由于P-N键化学稳定性差,导致其在酸和热条件下不稳定。而目前依赖酸性条件的O-磷酸化富集方法难以适用N-磷酸化富集,导致蛋白质N-磷酸化生物功能研究严重滞后。因此,迫切需要发展针对蛋白质N-磷酸化的高效富集方法。本文首先介绍了蛋白质N-磷酸化的结构特征和已报道的生物学功能,重点综述并分析了近20年来蛋白质N-磷酸化修饰富集方法,并对每一种富集方法的优缺点进行了评述,最后对潜在的富集方法进行了展望。Protein phosphorylation is one of the most common and important post-translational modifications that regulates almost all life processes.In particular,protein phosphorylation regulates the development of major diseases such as tumors,neurodegenerative diseases,and diabetes.For example,excessive phosphorylation of Tau protein can cause neurofibrillary tangles,leading to Alzheimer’s disease.Therefore,large-scale methods for identifying protein phosphorylation must be developed.Rapid developmentin efficient enrichment methods and biological mass spectrometry technologies have enabled the large-scale identification of low-abundance protein O-phosphorylation modifications in,allowing for a more thorough study of their biological functions.The N-phosphorylation modifications that occur on the side-chain amino groups of histidine,arginine,and lysine have recently received increased attention.For example,the biological function of histidine phosphorylation in prokaryotes has been well studied;this type of modification regulates signal transduction and sugar metabolism.Two mammalian pHis kinases(NME1 and NME2)and three pHis phosphatases(PHPT1,LHPP,and PGAM5)have been successfully identified using various biological methods.N-Phosphorylation is involved in multiple biological processes,and its functions cannot be ignored.However,N-phosphorylation is unstable under acidic and thermal conditions owing to the poor chemical stability of the P-N bond.Unfortunately,the current O-phosphorylation enrichment method,which relies on acidic conditions,is unsuitable for N-phosphorylation enrichment,resulting in a serious lag in the large-scale identification of protein N-phosphorylation.The lack of enrichment methods has also seriously hindered studies on the biological functions of N-phosphorylation.Therefore,the development of efficient enrichment methods that target protein N-phosphorylation is an urgent undertaking.Research on N-phosphorylation proteome enrichment methods is limited,hindering functional research.Thus,summarizing

关 键 词：蛋白质N-磷酸化 生物功能 富集方法 综述 

分 类 号：O658[理学—分析化学]