定量蛋白质组学分析ClpS在分枝杆菌耐药中的功能

Quantitative proteomics analysis of ClpS-mediated rifampicin resistance in Mycobacterium

摘要:

ClpS是原核生物蛋白质降解复合物ClpAPS的重要组成成分，它可以识别某些特定的氨基酸序列并将其呈递给ClpAP以促进其降解。同时，ClpS也抑制了其他蛋白质底物的降解。本研究通过在耻垢分枝杆菌中过度表达ClpS，发现所构建的重组菌株提高了利福平的抗药性。应用定量蛋白质组学技术，我们系统地分析了过度表达ClpS对于细菌蛋白质组的影响，并推测出细菌抗利福平的分子机制：ClpS促进稳态的调整、促进药物沉降以及加速药物代谢。本研究首次通过改变细菌降解复合物的相关蛋白的表达增加细菌的抗药性，并证明蛋白质组学技术是细菌的抗药性研究以及耐药株筛选的重要工具。

Abstract:

Adaptor protein ClpS is an essential regulator of prokaryotic ATP-dependent protease ClpAP, which delivers certain protein substrates with specific amino acid sequences to ClpAP for degradation. However, ClpS also functions as the inhibitor of the ClpAP-mediated protein degradation for other proteins. Here, we constructed the clpS-overexpression Mycobacterium smegmatis strain, and showed for the first time that overexpression of ClpS increased the resistance of M. smegmatis to rifampicin that is one of most widely used antibiotic drugs in treatment of tuberculosis. Using quantitative proteomic technology, we systematically analyzed effects of ClpS overexpression on changes in M. smegmatis proteome, and proposed that the increased rifampicin resistance was caused by ClpS-regulated drug sedimentation and drug metabolism. Our results indicate that the changes in degradation related proteins enhanced drug resistance and quantitative proteomic analysis is an important tool for understanding molecular mechanisms responsible for bacteria drug resistance.