广泛耐药结核分枝杆菌与标准菌株H37Rv菌体差异蛋白的比较

Analysis of Differential Proteins in Extensively Drug-resistant Mycobacterium Tuberculosis and H37Rv Standard Sensitive Strain

目的比较广泛耐药结核分枝杆菌临床分离株与标准菌株H37Rv菌体蛋白表达的差异,对差异蛋白质分类,以了解结核分枝杆菌广泛耐药可能的机制。方法通过双向凝胶电泳分离XDR菌株和H37Rv菌株菌体总蛋白,将蛋白表达图谱比较分析,差异蛋白点进行质谱鉴定。所获肽序列与生物信息数据库匹配,在NCBI及PIR数据库中查找蛋白质信息,并归纳分析。结果XDR菌株菌体蛋白表达图谱与H37Rv菌株存在差异,XDR菌株中有较多缺失及下调蛋白点,其中氧化还原酶类所占比例较大。切取77个差异点,获得53个蛋白点的肽质量指纹图谱和肽序列,鉴定出可作为潜在临床药物作用靶点的3个蛋白质:ATP依赖性假定蛋白Rv2623、半胱氨酸合酶和蛋白酶体。结论广泛耐药结核分枝杆菌临床分离株与标准株H37Rv在蛋白表达水平上存在差异。初步鉴定3个潜在临床药物作用靶点的蛋白质在进一步功能研究中,为抗广泛耐药结核分枝杆菌新药的研发提供依据。

Objective To observe and analysis differentially expressed protein spots （up-regula- ted, down-regulated, appearance and absence） of extensively drug-resistant Mycobaeterium tuberculo- sis clinical isolate and H37Rv standard sensitive strain, and classify differential proteins in order to un- derstand the mechanism of extepsively drug-resistant in Mycobacterium tuberculosis. Methods Proteins of two strains were separated into protein spots by two-dimensional gel electrophoresis and differenti- ally expressed spots analyzed by mass spectrometry. Peptide fingerprints and,its sequence searched in NCBI and PIR database for gene and protein information. Results There was an apparent difference in the images of two strains. 77 differential spots between extensively drug-resistant strain and H37Rv were discovered and cut out, and 53 Peptide fingerprints and sequences were obtained. Three proteins were identified as high-confidence drug targets in extensively drug-resistant strain： ATP -dependent hypothetical protein Rv2623, cysteine synthase and proteasome. Conclusion There is distinction be- tween extensively drug-resistant Mycobaeterium tuberculosis clinical isolate and H37Rv strain in pro- tein expressed level, which is helpful for understanding the mechanism and regulating information of drug resistance in Mycobacterium tuberculosis. The function analyses of three identified proteins are in the process, which may provide a basis of developing new drugs against drug-resistant Mycobacterium tuberculosis.