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

摘要:

[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iTRAQ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。

Abstract:

[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.