Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis, which mainly attacks the lungs and can lead to death in serious cases. It is one of the major public health problems of global concern. BDQ is an inhibitor targeting ATP synthetase of Mycobacterium tuberculosis, which can effectively inhibit the growth of Mycobacterium tuberculosis. It is the first new anti tuberculosis drug marketed in the global research and development of tuberculosis drugs for nearly half a century, and is also listed by the World Health Organization as the first drug of choice for rifampicin resistant tuberculosis and multi drug resistant tuberculosis long-term treatment schemes.

However, research has found that BDQ increases the risk of cardiac arrhythmia in patients due to its interaction with potassium channel protein hERG, and also has potential cross inhibitory activity on human ATP synthase. Therefore, revealing the working mechanism of Mycobacterium tuberculosis ATP synthase and the mechanism of action of BDQ, as well as the molecular mechanism of inhibiting human ATP synthase activity, is of great significance for the development of novel Mycobacterium tuberculosis ATP synthase inhibitors.

On July 3, 2024, Professor Gong Hongri and Academician Rao Zihe from the School of Life Sciences, Nankai University, together with Associate Researcher Liu Fengjiang from Guangzhou National Laboratory and Associate Researcher Gao Yan from the Institute of Immunochemistry, ShanghaiTech University, published a research paper titled "Inhibition of M. tuberculosis and human ATP synthase by BDQ and TBAJ-587" in Nature, explaining the molecular mechanism of Bedaquiline (BDQ) and its derivative TBAJ-587 in inhibiting Mycobacterium tuberculosis ATP synthase, and revealing their cross reaction mechanism with human ATP synthase, which has important guiding significance for the development of a new generation of highly selective anti tuberculosis drugs.


The research team first used the strategy of "gene knock in - gene knock out - gene overexpression" in combination with affinity chromatography and gel filtration chromatography protein purification methods to finally obtain a uniform, stable and active sample of Mycobacterium tuberculosis ATP synthase protein with the help of Mycobacterium smegmatis. Subsequently, the research team explored and optimized the preparation conditions of frozen samples, and successfully resolved the high-resolution cryo electron microscopy structure of Mycobacterium tuberculosis ATP synthase in BDQ bound state using single particle cryo electron microscopy technology (as shown in Figure 1). Research has found that BDQ mainly interacts strongly with Mycobacterium tuberculosis ATP synthase through quinoline groups (A group) and dimethylamino groups (D group) and binds to multiple sites in the transmembrane region, preventing the rotation of the c-ring of ATP synthase in the transmembrane region, thereby blocking proton transport and ultimately preventing ATP synthesis, achieving the goal of "starving" Mycobacterium tuberculosis.


Figure 1 Cryo electron microscopy structure of ATP synthase binding to BDQ in Mycobacterium tuberculosis

The most representative BDQ derivatives are TBAJ-587 and TBAJ-876, and currently the candidate drugs have entered clinical trials. The research team analyzed the high-resolution cryo electron microscopy structure of ATP synthase in Mycobacterium tuberculosis in the binding state of TBAJ-587 (as shown in Figure 2). The structure shows that the binding mode of TBAJ-587 to Mycobacterium tuberculosis ATP synthase is the same as that of BDQ. Moreover, TBAJ-587 and BDQ mainly interact with Mycobacterium tuberculosis ATP synthase through the A and D groups.


Figure 2 Cryo electron microscopy structure of Mycobacterium tuberculosis ATP synthase binding to TBAJ-587

Finally, the researchers found that both BDQ and TBAJ-587 exhibit cross reactivity towards human ATP synthase. Subsequently, the researchers successfully resolved the cryo electron microscopy structure of human ATP synthase binding to BDQ (as shown in Figure 3). Analysis found that TBAJ-587, which was redesigned based on the B and C groups in BDQ, only reduced the risk of cardiac arrhythmia caused by interaction with hERG protein. Only by redesigning and optimizing the A group can the interaction with human ATP synthase be reduced, thereby avoiding potential health risks in clinical treatment.


Figure 3 Cryo electron microscopy structure of human ATP synthase binding to BDQ

It is worth mentioning that the journal Nature also invited Professor Gregory Cook, former chairman of the Gordon Conference on Bioenergy, and his colleagues to write a highlight review titled "Enzyme blueprints will aid tuberculosis drug design" for this study in the News&Views column.

Zhang Yuying and Lai Yuezheng, doctoral students at Nankai University, are co first authors of this article. Professor Gong Hongri and Academician Rao Zihe from the School of Life Sciences, Nankai University, Associate Researcher Liu Fengjiang from Guangzhou National Laboratory, and Associate Researcher Gao Yan from the Institute of Immunochemistry, ShanghaiTech University are co corresponding authors.

The above work has been supported by projects such as the National Key Research and Development Program for Young Scientists, the National Excellent Youth Science Fund, and the China Association for Science and Technology Youth Talent Support Project. Researchers Peng Wei and Guo Yu from Guangzhou National Laboratory provided crucial support for this work. The imaging sub platform of Guangzhou National Laboratory and the biological electron microscopy platform of ShanghaiTech University have provided important technical support.

Original title: "Nature | Guangzhou National Laboratory and collaborators reveal the mechanism of action of the anti tuberculosis drug bedaquiline and its derivatives"

Read the original text

important clause
This article is uploaded and published by the author or organization of the Pengpai account on Pengpai News. It only represents the views of the author or organization and does not represent the views or positions of Pengpai News. Pengpai News only provides an information publishing platform.