采用MS-Fit进行肽质量指纹谱鉴定蛋白质时主要参数的优化  

Optimization of the major parameters of MS-Fit in peptide mass fingerprint identify proteins

作　　者：彭咏波[1] 母昭德[1] 马永平[2] 夏永鹏[1] 邱宗荫[1] 

机构地区：[1]重庆医科大学药学院药物分析教研室,重庆400016 [2]重庆医科大学生物化学与分子生物学教研室,重庆400016

出　　处：《重庆医科大学学报》2008年第7期769-772,787,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30476021);教育部博士点基金资助项目(教特发中心函[2004]166号)

摘　　要：目的:为了优化肽质量指纹谱(Peptide mass fingerprinting,PMF)鉴定蛋白质时采用MS-Fit引擎搜索的分析参数。方法:将2-DE分离后的Carbonic anhydrase-2和BSA进行胶内充分酶解,肽段经过MALDI-TOF-MS分析得到PMF数据。选择Swissprot数据库,以Carbonic anhydrase2为模型优化搜索参数。结果:主要搜索参数的最佳设置为:半胱氨酸修饰为Carbamidomethylation,肽质量容错模型为百分数,在研究中0.1%容错数为最好。结论:本文通过标准蛋白对搜索主要参数的优化,建立了方便、可靠的MS-Fit搜索参数模式。Objective：To optimize the major parameters of bioinformatics analysis conditions in process of PMF（Peptide Mass Fingerprinting） identify target proteins. Methods：In this process of experiment design and analysis,we made the Bovine Carbonic anhydrase-2 and BSA from 2-DE gel,after proteolysis with porcine trypsin and gathered the peptides,subsequently identified with MALDI-TOF MS and got the peptides mass data. Then,we selected database of Swissprot and MS-Fit search engine,the standard Carbonic anhydrase-2 protein used as model to set up protein search standard conditions. Results：Our study showed that the cysteine was modified by carbamidomethylation more convenient in protein search,at the same time,the better mode of mass tolerance was percentage and 0.1% was suitable tolerance in our search. Then we used BSA mass data to check the accuracy of MS search standard conditions. Conclusions： Results show that the optimized parameters are reliable.

关 键 词：MALDI—TOF—MS PMF搜索 蛋白质 MS—Fit 

分 类 号：R917[医药卫生—药物分析学]