利用双向电泳技术研究异烟肼耐药MTB感染U937引致的差异表达蛋白  

Analysis of Differential Expression Proteins of U937 Infected with Isoniazid-resistant MTB by Two-dimensional Electrophoresis

作　　者：刘丽蓉[1] 乐军[1] 王洪海[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《中国生物化学与分子生物学报》2006年第2期142-150,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展规划(973计划)(No.2002CB512804)项目;国家教育部博士点基金资助项目(No.20020246019)~~

摘　　要：利用双向电泳技术对结核分枝杆菌(MTB)异烟肼(INH)耐药株和敏感株感染人源巨噬细胞(U937)后的全细胞蛋白表达图谱进行差异比较和分析,发现其中产生差异的有86个蛋白质斑点.利用基质辅助激光解吸/电离飞行时间质谱技术对其中8个差异表达蛋白质斑点进行分析鉴定,获得8个明确的肽质量指纹图谱.通过在蛋白质数据库中进行检索分析,确定这8个蛋白质中的2个为在INH耐药株感染的U937中差异表达的干细胞生长因子和主要组织相容性复合物Ⅰ类抗原,6个为在敏感株感染的U937中差异表达的微管蛋白4β、信号转导和转录激活子3、延长因子-2激酶、环指蛋白29、锌指蛋白193和SNARE Vti1a-β蛋白.实验结果显示,INH耐药株和敏感株感染后的U937表达蛋白有差异,这有助于分析解释临床中观察到的受INH耐药株感染的病例出现毒力下降、致病性下降、传染性降低以及潜伏期延长的现象.结果为针对INH耐药株进行的新疫苗的设计提供了探索方向.Two-dimensional electrophoresis （2-DE） were employed to compare the global protein patterns between human macrophage cell line U937 infected with Mycobacterium Tuberculosis （MTB） isoniazid（INH）- resistant strains and INH-susceptible strains, 86 protein spots were expressed differentially. Eight differential expression protein spots were measured by matrix assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. The data obtained from peptide mass fingerprinting were used for protein database search. Two protein spots that differentially express in U937 infected with MTB INH-resistant strains were identified as stem cell growth factor, MHC class Ⅰ antigen, respectively. Six protein spots that differentially express in U937 infected with MTB INH-susceptible strains were identified as tubulin β4, signal transducer and activator of transcription 3 （STAT 3）, elongation factor-2 kinase （eEF-2K）, ring finger protein 29, zinc finger protein 193 and SNARE Vtila-β protein. These data provide insight into the changed global protein patterns of the U937 infected with MTB INH-resistant strains and INH-susceptible strains, and interpret the lower virulence, lower pathogenicity, lower infectivity, and longer incubation period that infected with INH-resistant strains in clinical cases. These results may contribute to the further research of new vaccines against INH-resistant strains.

关 键 词：结核分枝杆菌 异烟肼 耐药 巨噬细胞 双向电泳 差异表达 

分 类 号：R52[医药卫生—内科学]