Identification of the inhibitory mechanism of ecumicin and rufomycin 4-7 on the proteolytic activity of Mycobacterium tuberculosis ClpC1/ClpP1/ClpP2 complex
journal contributionposted on 2023-03-14, 21:19 authored by Jeongpyo Hong, Nguyen Minh Duc, Byeong-Chul Jeong, Sang Hyun ChoSang Hyun Cho, Gauri Shetye, Jin Cao, Cherlhyun Jeong, Hanki Lee, Joo-Won Suh
Ecumicin and rufomycin 4-7 disrupt protein homeostasis in Mycobacterium tuberculosis by inhibiting the proteolytic 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 proteolytic 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 population 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.
“Targeting Protein Degradation ClpC1 ATPase” | Funder: National Institutes of Health | Grant ID: U19AI142735
CitationHong, J., Duc, N. M., Jeong, B. C., Cho, S., Shetye, G., Cao, J., Lee, H., Jeong, C., Lee, H.Suh, J. W. (2023). Identification of the inhibitory mechanism of ecumicin and rufomycin 4-7 on the proteolytic activity of Mycobacterium tuberculosis ClpC1/ClpP1/ClpP2 complex. Tuberculosis, 138, 102298-. https://doi.org/10.1016/j.tube.2022.102298
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3207 Medical Microbiology32 Biomedical and Clinical Sciences3202 Clinical Sciences3204 ImmunologyRare DiseasesTuberculosis3 Good Health and Well BeingMycobacterium tuberculosisBacterial ProteinsAdenosine TriphosphatasesAbnormal ClpC1ClpC1EcumicinIntermolecular dynamics of ClpC1Rufomycin 4-7Microbiology3202 Clinical sciences3207 Medical microbiologyMedical and Health Sciences