基于电子转移/高能碰撞解离质谱碎裂的O-GlcNAc糖基化位点定量分析方法在高脂喂养小鼠肝脏糖蛋白质组分析中的应用  

O-GlcNAcylation Quantification Based on Electron-Transfer/Higher-Energy Collisional Dissociation and Its Application in O-GlcNAcylation Site Mapping of Liver Proteomics in High Fat Diet-fed Mice

作　　者：邵文亚 梁玉[2] 刘键熙[4] 刘洪涛[3] 王钊伟[5] 随志刚 赵宝锋[2] 张晓丹 梁振[2] 张丽华[2] 张玉奎[2] SHAO Wen-Ya;LIANG Yu;LIU Jian-Xi;LIU Hong-Tao;WANG Zhao-Wei;SUI Zhi-Gang;ZHAO Bao-Feng;ZHANG Xiao-Dan;LIANG Zhen;ZHANG Li-Hua;ZHANG Yu-Kui(Department of Preventive Medicine,School of Public Health,Fujian Medical University,Fuzhou 350122,China;CAS Key Lab of Separation Science for Analytical Chemistry,National Chromatographic Research and Analysis Center,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,China;State Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,China;College of Environmental Science and Engineering,Fujian Normal University,Fuzhou 350007,China;Institute of Computing Technology,Chinese Academy of Sciences,Beijing 100190,China)

机构地区：[1]福建医科大学公共卫生学院预防医学系,福州350122 [2]中国科学院大连化学物理研究所,中国科学院分离分析化学重点实验室,国家色谱研究分析中心,大连116023 [3]中国科学院过程工程研究所,生化工程国家重点实验室,北京100190 [4]福建师范大学环境科学与工程学院,福州350007 [5]中国科学院计算技术研究所,北京100190

出　　处：《分析化学》2021年第12期2106-2116,I0030-I0036,共18页Chinese Journal of Analytical Chemistry

基　　金：福建省自然科学基金(No.2020J01641);国家自然科学基金(No.21605020);福建医科大学基金项目(No.2018QH1007)资助。

摘　　要：O-连接N-乙酰葡糖胺翻译后修饰(O-GlcNAc)是发生在蛋白质丝氨酸或者苏氨酸位点上的单糖修饰,由于其具有化学计量数少、在质谱鉴定中离子化效率低以及无特定的氨基酸序列等特点,使O-GlcNAc翻译后修饰位点的定性和定量分析难度较大。本研究结合基于凝集素弱亲和色谱的O-GlcNAc修饰糖肽富集方法、本研究组发展的准等重六重标记定量方法和电子转移/高能碰撞解离(EThcD)模式的高分辨质谱技术,发展了一种O-GlcNAc修饰位点的高通量定量分析方法,对高脂喂养小鼠肝脏中蛋白质O-GlcNAc修饰位点进行规模化定量分析。共定量分析了783个O-GlcNAc位点,其中122个位点的表达量存在明显差异,对应于85个O-GlcNAc蛋白,并初步探讨了脂代谢过程中O-GlcNAc修饰的作用。本研究有望为肥胖和胆固醇酯沉积等营养代谢相关疾病提供新的治疗思路。O-linkedβ-N-acetylglucosaminylation(O-GlcNAcylation)is a single carbohydrate moiety post-translational modification that occurs on serine and threonine side chains of intracellular protein and plays an important role in nutrient metabolism.However,the great research challenges are the inherently low stoichiometry,poor ionization efficiency of O-GlcNAc peptides and no specific animo acid sequence.In this study,a strategy for quantitative detection of O-GlcNAc sites was developed by combining pseudo-isobaric dimethyl strategy,which was applied for the O-GlcNAcylation sites profiling towards the mice liver.In total,783 O-GlcNAc sites were unambiguously quantified from high-fat fed mice liver.Among which,122 O-GlcNAc sites were differentially expressed,corresponding to 85 O-GlcNAc proteins.Finally,the biological function of the differentially expressed O-GlcNAc proteins was analyzed.

关 键 词：O-连接N-乙酰葡糖胺修饰 定量蛋白质组学 电子转移/高能碰撞解离 

分 类 号：R341[医药卫生—基础医学] O657.63[理学—分析化学]