【摘要】 利用双向电泳技术,对猫爪草提取物作用前后的结核分枝杆菌临床分离株的全细胞蛋白表达图谱进行差异比较和分析,发现其中22个蛋白质斑点的浓度具有差异,利用基质辅助激光解吸/电离飞行时间质谱技术,对其中4个表达明显下调和1个明显上调的蛋白质斑点进行分析鉴定,获得5个明确的肽质量指纹图谱.通过数据库检索,确定这5个蛋白质分别为S-腺苷甲硫氨酸合成酶、吲哚-3-甘油磷酸合酶、烯酰-CoA水合酶、琥珀酰辅酶A合成酶和60 kD的分子伴侣2.其中前4个分子是首次报道参与结核分枝杆菌的重要生理活动.该结果有助于了解猫爪草提取物对结核分枝杆菌生理的影响,为进一步确定中药猫爪草提取物对结核分枝杆菌的作用靶标和机理提供了基础.更多还原

【Abstract】 Radix Ranuncoli Ternati is clinically effective traditional Chinese medicine for multidrug resistant tuberculosis.Its active components and mechanism of action remain unsolved.Two dimensional electrophoresis(2-DE)was employed to address this problem.Globlal proteins between Mycobacterium tuberculosis untreated and that treated with Radix Ranuncoli Ternati were compared,and 22 protein spots were found to be expressed differentially.The 4 protein spots which remarkably decreased and 1 spot upregulated in Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati were subjected to matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI TOF MS) analyze.The data obtained from peptide mass finger printing were used in protein data base search.The 4 protein spots in gel were identified as metK(S-adenosylmethionine synthetase),trpC(indole-3-glycerol phosphate synthase),echA8(enoyl-CoA hydratase),sucD(succinyl-CoA synthase) and the up-regulated spot as groEL2(60 kD chaperonin 2),which are involved in amino acid and fatty acid metabolism and TCA cycle,respectively.These data not only revealed the changed global protein patterns of Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati extracts,but also provide potential novel anti-tuberculosis drug targets.更多还原