Bundutidi Gloria Mavinga, Mochizuki Kota, Matsuo Yuichi, Hayashishita Mizuki, Sakura Takaya, Ando Yuri, Cook Gregory Murray, Rajib Acharjee, Bringaud Frédéric, Boshart Michael, Hamano Shinjiro, Sekijima Masakazu, Hirayama Kenji, Kita Kiyoshi, Inaoka Daniel Ken
Program for Nurturing Global Leaders in Tropical and Emerging Communicable Disease, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Department of Parasitology, Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.
Commun Biol. 2025 Feb 6;8(1):187. doi: 10.1038/s42003-025-07611-0.
The F-type ATP synthase inhibitor bedaquiline (BDQ) is a potent inhibitor of mycobacterial growth and this inhibition cannot be rescued by fermentable carbon sources that would supply ATP by an alternative pathway (substrate level phosphorylation). To gain mechanistic insight into this phenomenon, we employed a metabolic engineering approach. We introduced into Mycobacterium smegmatis an alternative ATP production pathway by substrate-level phosphorylation, specifically through overexpression of trypanosomal acetate:succinate co-enzyme A (CoA) transferase (ASCT). Intriguingly, the overexpression of ASCT partially restored intracellular ATP levels and resulted in acquired tolerance to BDQ growth inhibition at low, but not high concentrations of BDQ. These results implicate intracellular ATP levels in modulating the growth inhibitory activity of BDQ at low concentrations. These findings shed light on the intricate interplay between BDQ and mycobacterial energy metabolism, while also providing a novel tool for the development of next-generation ATP synthase-specific inhibitors targeting mycobacteria.
F型ATP合酶抑制剂贝达喹啉（BDQ）是分枝杆菌生长的强效抑制剂，这种抑制作用无法通过可发酵碳源来挽救，因为这些碳源会通过替代途径（底物水平磷酸化）提供ATP。为了深入了解这一现象的机制，我们采用了代谢工程方法。我们通过底物水平磷酸化，特别是通过过表达锥虫乙酸：琥珀酸辅酶A（CoA）转移酶（ASCT），在耻垢分枝杆菌中引入了一条替代的ATP产生途径。有趣的是，ASCT的过表达部分恢复了细胞内ATP水平，并导致在低浓度而非高浓度的BDQ下获得了对BDQ生长抑制的耐受性。这些结果表明细胞内ATP水平在调节低浓度BDQ的生长抑制活性中起作用。这些发现揭示了BDQ与分枝杆菌能量代谢之间复杂的相互作用，同时也为开发针对分枝杆菌的下一代ATP合酶特异性抑制剂提供了一种新工具。
