Hong, Jeongpyo Duc, Nguyen Minh Jeong, Byeong-Chul Cho, Sanghyun Shetye, Gauri Cao, Jin Lee, Hyun Jeong, Cherlhyun Lee, Hanki Suh, Joo-Won 2024-01-19T10:30:41Z 2024-01-19T10:30:41Z 2023-02-10 2023-01 1472-9792 https://pubs.kist.re.kr/handle/201004/114128 Ecumicin and rufomycin 4-7 disrupt protein homeostasis in Mycobacterium tuberculosis by inhibiting the pro-teolytic activity of the ClpC1/ClpP1/ClpP2 complex. Although these compounds target ClpC1, their effects on the ATPase activity of ClpC1 and proteolytic activity of ClpC1/ClpP1/ClpP2 vary. Herein, we explored the ClpC1 molecular dynamics with these compounds through fluorescence correlation spectroscopy. The effect of these compounds on the ATPase activity of ClpC1-cys, the recombinant protein for fluorescence labeling, and pro-teolytic activity of ClpC1-cys/ClpP1/ClpP2 were identical to those of native ClpC1, whereas the intermolecular dynamics of fluorescence-labelled ClpC1 were different. Treatment with up to 1 nM ecumicin increased the population of slower diffused ClpC1 components compared with ClpC1 without ecumicin. However, this pop-ulation was considerably reduced when treated with 10 nM ecumicin. Rufomycin 4-7 treatment resulted in a slower diffused component of ClpC1, and the portion of this component increased in a concentration-dependent manner. Ecumicin can generate an abnormal ClpC1 component, which cannot form normal ClpC1/ClpP1/ClpP2, via two different modes. Rufomycin 4-7 only generates slower diffused ClpC1 component that is inadequate to form normal ClpC1/ClpP1/ClpP2. Overall, we demonstrate that ecumicin and rufomycin 4-7 use different action mechanisms to generate abnormal ClpC1 components that cannot couple with ClpP1/ClpP2. English Churchill Livingstone Identification of the inhibitory mechanism of ecumicin and rufomycin 4-7 on the proteolytic activity of Mycobacterium tuberculosis ClpC1/ClpP1/ ClpP2 complex Article 10.1016/j.tube.2022.102298 1 Tuberculosis, v.138 Tuberculosis 138 Y scie scopus 000918637900001 2-s2.0-85144984290 Immunology Microbiology Respiratory System Immunology Microbiology Respiratory System Article Ecumicin Rufomycin 4-7 ClpC1 Intermolecular dynamics of ClpC1 Abnormal ClpC1