Dynamic Proteomic and PTMomic Characterization of Mycobacteria after Clinical Pharmaceutical Intervention

Tuberculosis, a major global health threat, necessitates understanding the pharmacological mechanisms of current drugs to combat multidrug resistance. In addition to alterations at the proteome level, the dynamic changes occurring at various levels of post-translational modifications (PTMs) following pharmacological intervention remain unclear. In our current study, we employed a quantitative proteomic approach to systematically analyze the dynamic molecular alterations at both the proteome and PTM levels in response to clinical drugs, including ethambutol, bedaquiline, moxifloxacin, and streptomycin. Our findings revealed enriched bioprocesses beyond known functions, phosphorylation-level changes in kinases and phosphatases, and increased acetylation levels with all four drugs. Overexpression of CobB in Mycobacterium smegmatis significantly increased its susceptibility to ethambutol, indicating enhanced drug sensitivity. Our study provides integrated multiomics resources for understanding the dynamic molecular characteristics and drug resistance associated with clinical drug interventions and proposes novel therapeutic strategies targeting the PTM levels.