基于化学衍生和CID碎裂模式鉴定蛋白精氨酸-ADP-核糖基化的新方法  

Novel Method for Identification of Protein Arginine-ADP-ribosylation Based on Chemical Derivatization and CID Fragmentation

作　　者：赵志强[1,2] 邱辉华[1] 姚军[1,2] 陈永峰[1] 

机构地区：[1]台州学院医学院 [2]肿瘤研究所,台州318000

出　　处：《高等学校化学学报》2013年第12期2704-2709,共6页Chemical Journal of Chinese Universities

基　　金：浙江省自然科学基金(批准号:LY12C05002)资助

摘　　要：建立了一种新的基于碰撞诱导解离(CID)碎裂模式鉴定精氨酸-腺苷二磷酸(ADP)-核糖基化多肽的新方法.首先,在碱性条件下将精氨酸-ADP-核糖基化血管紧张素-Ⅰ转变为鸟氨酸化血管紧张素-Ⅰ,或在磷酸二酯酶和碱性磷酸酶处理下水解为精氨酸核糖基化血管紧张素-Ⅰ,然后对上述2种衍生物进行基于CID碎裂模式的串联质谱分析.结果表明,与未衍生的精氨酸-ADP-核糖基化血管紧张素-Ⅰ相比,在鸟氨酸化血管紧张素-Ⅰ和精氨酸核糖基化血管紧张素-Ⅰ的质谱图上发现大部分来自于肽骨架碎裂的离子峰,可提供足够的序列信息以确定精氨酸-ADP-核糖基化位点.High performance liquid chromatography coupled to electrospray ionization mass spectrometry（MS） was widely used to map protein posttranslational modifications in complex biological samples. But frequently- used collision induced dissociation （CID） was unsuccessful in fragmentizing arginine adenosine diphosphate （ADP）-ribosylated peptides because the spectra from CID fragmentation were dominated by ADP-ribose frag- ment ions at the expense of the fragments generated from peptide backbone, making the assignment of the pep- tide sequence very difficult. In this work, a novel method was developed for identification of arginine ADP-ri- bosylated peptides based on CID fragmentation. Arginine ADP-ribosylated angiotensin-I was either converted to ornithinylated angiotensin-I under alkaline condition or hydrolyzed to arginine ribosylated angiotensin- I by treatment with phosphodiesterase and alkaline phosphatase. The two derivatives were then subjected to CID fragmentation during tandem MS analysis. Compared with unmodified ADP-ribosylated angiotensin-1 derivatives generated more sequence specific ions in the spectra, giving enough information to localize of arginine ADP-ribosylation. the two the site

关 键 词：血管紧张素-I 化学衍生 碰撞诱导解离 质谱 精氨酸 ADP-核糖基化 

分 类 号：O657.63[理学—分析化学]