Gao Yamin, Fang Cuiting, Zhou Biao, Hameed H M Adnan, Sun Changli, Tian Xirong, He Jing, Han Xingli, Zhang Han, Li Jun, Ju Jianhua, Chen Xinwen, Zhong Nanshan, Ma Junying, Xiong Xiaoli, Zhang Tianyu
State Key Laboratory of Respiratory Disease, Institute of Drug Discovery, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
Guangdong-HongKong-Macau Joint Laboratory of Infectious Respiratory Diseases, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
Commun Biol. 2025 Aug 13;8(1):1219. doi: 10.1038/s42003-025-08646-z.
The mycobacterial caseinolytic protease (Clp) system has been recognized as a promising therapeutic target. In this study, we identify two novel ilamycin analogs, ilamycin E (ILE) and ilamycin F (ILF), both targeting the ClpC1 component of the ClpC1P1P2 proteasome. ILE potently disrupts ClpC1P1P2-mediated proteolysis, leading to delayed bactericidal activity, while ILF also binds ClpC1, albeit with lower affinity. Notably, we discover and validate a unique mutation in clpX and a novel insertion in clpC1 both conferring resistance to ILE and ILF in mycobacterium by gene editing. Furthermore, ILE can also inhibit the proteolytic activity of ClpXP1P2 in a manner dependent on the substrate's tag sequence and adaptor. This first demonstration of clpX- and clpC1-mediated ilamycins resistance underscores the potential of ilamycins to target multiple components of the Clp protease system, offering a novel dual-target strategy for combating mycobacterial infections.
分枝杆菌酪蛋白水解蛋白酶（Clp）系统已被认为是一个有前景的治疗靶点。在本研究中，我们鉴定出两种新型伊拉霉素类似物，即伊拉霉素E（ILE）和伊拉霉素F（ILF），二者均靶向ClpC1P1P2蛋白酶体的ClpC1组分。ILE能有效破坏ClpC1P1P2介导的蛋白水解作用，导致杀菌活性延迟，而ILF也能结合ClpC1，尽管亲和力较低。值得注意的是，我们通过基因编辑发现并验证了clpX中的一个独特突变和clpC1中的一个新型插入，二者均赋予分枝杆菌对ILE和ILF的抗性。此外，ILE还能以依赖于底物标签序列和衔接子的方式抑制ClpXP1P2的蛋白水解活性。clpX和clpC1介导的伊拉霉素抗性的首次证明强调了伊拉霉素靶向Clp蛋白酶系统多个组分的潜力，为对抗分枝杆菌感染提供了一种新型双靶点策略。
