纳升电喷雾串联质谱鉴定吗啡依赖2个重要相关蛋白的N端乙酰化修饰  

Characterization of N-terminal acetylation of two morphine dependence related proteins with nanoESI-MS/MS

作　　者：何家田[1] 刘炳玉[1] 吴宁[2] 苏瑞斌[2] 李锦[2] 张学敏[1] 魏开华[1] 王红霞[1] 

机构地区：[1]国家生物医学分析中心,北京100850 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《药物分析杂志》2008年第2期171-175,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家863计划资助项目(No.2002AA232021);973计划资助项目(2003CB515400)

摘　　要：目的:用纳升电喷雾串联质谱(nanoESI-MS/MS)鉴定吗啡依赖相关蛋白。方法:采用吗啡递增给药法建立吗啡依赖模型,用纳洛酮催促后出现典型的戒断症状,以盐水组为对照,对长期接受吗啡处理和纳洛酮催促戒断这2种状态的大鼠纹状体进行了比较蛋白质组研究,确定了33个差异点。其中差异点150和209用电喷雾串联质谱进行了氨基酸序列分析。结果:电喷雾串联质谱分析表明差异点150和209为2个重要的吗啡依赖蛋白质:G 蛋白β1亚基和电压依赖型阴离子通道蛋白1(VDAC-1),并且这2个蛋白质都发生了 N 端乙酰化修饰。结论:纳升电喷雾串联质谱分析是研究蛋白质 N 端乙酰化的好方法。N 端乙酰化对蛋白质生物学功能有重要作用。这2个蛋白质 N 端乙酰化的生物学功能及其与吗啡依赖的关系值得深入研究。Objective：To characterize morphine dependence differential proteins by nano electrospray tandem mass spectrometry （ nanoESI - MS/MS）. Method： The rats were treated with ascending doses of morphine to achieve dependence and leading to characteristic withdrawal syndromes after naloxone - precipitated withdrawal. Comparative proteomic analysis was performed in striatum of two phases of morphine dependence, long - term exposure to morphine and drug withdrawal and 33 different protein spots were identified by matrix - assisted laser desorption/ionization - time - of - flight - mass spectrometry （ MALDI - TOF - MS）. The differential spot 150 and 209 were analyzed by nanoESI - MS/MS since they were not identified by MALDI - TOF - MS unambiguously. Results： NanoESI - MS/MS analysis indicated that spot 150 and 209 were G proteinβ1 subunit and voltage- dependent anion -selective channel protein 1 （ VDAC - 1 ） respectively. Most importantly, nanoESI - MS/MS analysis showed that the two proteins are all N - terminally acetylated. Conclusion： NanoESI - MS/MS is a powerful tool for N - terminal acetylation analysis. N - terminal acetylation is critical to biological function of proteins. It is worth of giving insight into function of the two N - terminal acetylated proteins and their relations to morphine dependence.

关 键 词：吗啡依赖 乙酰化 G蛋白|B1亚基 电压依赖型阴离子通道蛋白1(VDAC-1) 纳升电喷雾串联质谱(nanoESI-MS/MS 

分 类 号：R917[医药卫生—药物分析学]