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

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

【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。

[Objective] To investigate the differentially expressed proteins between isoniazid,streptomycin monoand 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 H37 Rv.Differentially expressed proteins were identified through isobaric tags for relative and absolute quantification（i TRAQ） 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 H37 Rv.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（Rv2986 c and Rv1908 c） were up-regulated over 1.25 folds and two proteins（Rv3133 c 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 Rv2986 c,Rv1908 c,Rv3133 c and Rv0577 is likely to be related to isoniazid and streptomycin resistance in M.tuberculosis.