结核分枝杆菌耐药株和敏感株菌体碱性蛋白的比较蛋白质组学研究

Comparison of the proteome of Mycobacterium tuberculosis cytosolic basic proteins between multiple drug-resistant stains and sensitive stains

目的研究结核分枝杆菌耐多药菌株与敏感菌株菌体碱性蛋白的蛋白质表达差异。方法运用双向电泳技术(2-DE)比较8株耐多药菌株与9株敏感菌株的菌体碱性蛋白表达图谱差异,差异蛋白进行胶内酶切,肽混合物使用基质辅助激光解吸-电离飞行时间质谱仪(MALDI-TOF-MS)进行质谱分析,将肽质量指纹谱数据输入互联网上的蛋白质数据库进行检索。结果比较发现重复性较好的差异蛋白质斑点10个,获得5个明确的肽质量指纹谱,鉴定5个蛋白分别为:NADH-CoQ氧化还原酶B链(NuoB,Rv3146)、乙酰羟酸合成酶小亚基(IlvN,Rv3002c)以及3个假定蛋白,分别由Rv2159c、Rv0068、Rv0992编码。结论上述蛋白的鉴定在深入研究结核分枝杆菌耐药形成机制,筛选具有潜在价值的结核药物靶位和耐多药结核病诊断的分子标志物方面具有参考价值。

Objective To study cytesolic basic protein variation between multiple drug-resistant M. tuberculosis and sentisitive M. tuberculosis. Methods Two-dimensional electrophoresis （2-DE） were employed to compare the differential expression proteins between 8 multiple drug-resistant strains and 9 sentisitive strains. The differential proteins were digested in gel by enzyme and the mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. The data obtained from peptide mass fingerprinting （PMF） were searched using the intemet available database. Results There were ten re- peated differential spots were found and six of them were identified in gel as namely, hypothetical protein Rv2159e, hypothetical protein Rv0068, NuoB, llvN, hypothetical protein Rv0992. Conclusion These proteins are potentially target sites for chemotherapy against M. tuberculosis and new marker in the detection of resistance of M. tuberculosis, at the same time, this study provides further insights into the mechanisms of drug-resistance.