相对定量分析异烟肼、链霉素单耐药与多耐药结核分枝杆菌临床分离株的蛋白质组差异

【目的】探讨异烟肼(isoniazid,INH)、链霉素(streptomycin,SM)单耐药结核分枝杆菌(Mycobacterium tuberculosis,MTB)与INH/SM多耐药MTB蛋白质组差异。【方法】应用i TRAQ结合Nano LC-MS/MS定量蛋白质组学技术,分析临床分离INH、SM或INH/SM耐药MTB与H37Rv标准株间均表达差异蛋白;并以INH/SM耐药MTB与H37Rv比值为对照,相对定量分析单耐药与多耐药MTB蛋白表达差异倍数;运用DAVID 6.7分析差异蛋白生物功能;STITCH 5.0分析差异蛋白与INH和SM相互作用。【结果】与H37Rv标准株比较,58个蛋白在INH、SM耐药与INH/SM耐药MTB间均有表达差异,共同差异蛋白生物功能主要为氧化还原酶活性和转移酶活性;主要参与丙酸代谢信号通路。共同差异蛋白中,与INH/SM耐药MTB比较,Rv2986c和Rv1908c在INH、SM耐药MTB均表达上调1.25倍;Rv3133c和Rv0577则均表达下调0.7倍;生物信息学预测发现以上4种蛋白可直接或间接与INH、SM进行相互作用。【结论】INH、SM单耐药和INH/SM多耐药MTB蛋白表达谱有较大差异,蛋白Rv2986c、Rv1908c、Rv3133c和Rv0577表达水平及相互作用可能与INH和SM耐药有关。

Quantitative proteomics analysis of isoniazid, streptomycin mono-drug resistant and poly-drug resistant clinical isolates of Mycobacterium tuberculosis

Objective] To investigate the differentially expressed proteins between isoniazid, streptomycin mono-and poly-drug resistant clinical isolate strains of Mycobacterium tuberculosis. Methods] Cellular proteins were extracted from isoniazid, streptomycin mono-and poly-drug resistant clinical isolates M. tuberculosis and H37Rv. Differentially expressed proteins were identified through isobaric tags for relative and absolute quantification (iTRAQ) combined with Nano LC-MS/MS technology. The biological function and interaction among differentially expressed proteins and isoniazid or streptomycin were analyzed by DAVID 6.7 and STITCH 5.0, respectively. Results] 58 differentially expressed proteins were identified in isoniazid, streptomycin mono-drug resistant strains and isoniazid, streptomycin poly-drug resistance strain compared with the proteomic profiles of H37Rv. The biological function of differentially expressed proteins mainly relates to oxidoreductase and transferase activity. Compared with isoniazid and streptomycin poly-drug resistance strain, two proteins (Rv2986c and Rv1908c) were up-regulated over 1.25 folds and two proteins (Rv3133c and Rv0577) were down-regulated less than 0.7 fold in isoniazid, streptomycin mono-drug resistant strains. Bioinformatics predicted that the four proteins interact with isoniazid and streptomycin directly or indirectly. Conclusion] The expressed level or the interactions together of Rv2986c, Rv1908c, Rv3133c and Rv0577 is likely to be related to isoniazid and streptomycin resistance in M. tuberculosis.