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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.
Two-dimensional electrophoresis technique was used to compare and analyze the whole cell protein expression profiles of the clinical isolates of Mycobacterium tuberculosis before and after the action of the cat’s paw extract, and the concentration of 22 protein spots was found to be different, using matrix-assisted laser desorption/ Ionization time-of-flight mass spectrometry was used to analyze and identify four protein spots that were significantly down-regulated and one significantly up-regulated. Five clear peptide mass fingerprints were obtained. Through database search, these five proteins were identified as S-adenosine. Methionine synthase, indole-3-glycerophosphate synthase, enoyl-CoA hydratase, succinyl-CoA synthetase, and 60 kD chaperone 2. The first four of these are the first reported involvement of tuberculosis The important physiological activity of Bacillus. This result helps to understand the influence of the extract of cat’s claw on the physiology of Mycobacterium tuberculosis, and provides the basis for further determination of the target and mechanism of the action of the extract of Chinese herb catus madame on Mycobacterium tuberculosis.