蛋白质组研究中肽质量指纹谱鉴定方法的建立及应用  

Development and Preliminary Application of a Peptide Mass Fingerprinting Technique in Proteome Research

作　　者：王京兰[1] 万晶宏[2] 罗凌[2] 熊少祥[3] 王光辉[3] 贺福初[2] 钱小红[1] 

机构地区：[1]军事医学科学院国家生物医学分析中心,北京100850 [2]军事医学科学院放射医学研究所,北京100850 [3]中国科学院化学所国家质谱中心,北京100080

出　　处：《生物化学与生物物理学报》2000年第4期373-378,共6页

基　　金：国家自然科学基金!重大项目资助课题No .3 9990 60 0 ;军事医学科学院科技创新启动基金资助课题!( 1998~ 1999)&&

摘　　要：建立了用肽质量指纹谱和数据库检索方法鉴定凝胶电泳分离蛋白质的方法。用标准蛋白质对胶上蛋白质原位酶切制备肽谱的方法进行了讨论。分析了实际细胞蛋白质样品 ,获得了双向电泳分离的人肺癌细胞蛋白质谱中三个蛋白质点的肽指纹谱 ,并通过数据库检索分别鉴定为甘油醛 3 磷酸脱氢酶 2、遍在蛋白羧基末端水解同工酶和丙糖磷酸异构酶。Proteome means the total proteins expressed by the genome in a cell or tissue. Two dimensional electrophoresis (2 DE) is now the most powerful separating technique and the key separation method used in proteome. Peptide mass fingerprinting (PMF) is becoming a widely used and vastly develo ped technique for protein identification in 2 D gels. In this research, a systematic method to identify the proteins in polyacrylamide gels by PMF was develo ped. Proteins were digested in gel by enzyme and the masses of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI TOF MS). The data obtained from PMF were used in protein database search and the protein identification. The proteins from human lung cancer cells isolated by 2 DE were subjected to identification by the PMF method developed in this work. Three spots of proteins in gel were identified as G3P2 HUMAN, UBL1 HUMAN and TPIS HUMAN.

关 键 词：蛋白质组 肽质量指纹谱 MALDI-TDF-MS 肿瘤细胞 

分 类 号：Q51[生物学—生物化学]