Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption
To identify novel inhibitors of Mycobacterium tuberculosis cell envelope biosynthesis, we employed a two-step approach. First, we screened the diverse synthetic small molecule 71,544-compound Enamine library for growth inhibitors using the non-pathogenic surrogate Mycobacterium bovis BCG as screenin...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-12-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.02960/full |
| _version_ | 1818116386411511808 |
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| author | Annanya Shetty Annanya Shetty Zhujun Xu Umayal Lakshmanan Jeffrey Hill Meng Ling Choong Shu-Sin Chng Shu-Sin Chng Yoshiyuki Yamada Anders Poulsen Thomas Dick Thomas Dick Martin Gengenbacher |
| author_facet | Annanya Shetty Annanya Shetty Zhujun Xu Umayal Lakshmanan Jeffrey Hill Meng Ling Choong Shu-Sin Chng Shu-Sin Chng Yoshiyuki Yamada Anders Poulsen Thomas Dick Thomas Dick Martin Gengenbacher |
| author_sort | Annanya Shetty |
| collection | DOAJ |
| description | To identify novel inhibitors of Mycobacterium tuberculosis cell envelope biosynthesis, we employed a two-step approach. First, we screened the diverse synthetic small molecule 71,544-compound Enamine library for growth inhibitors using the non-pathogenic surrogate Mycobacterium bovis BCG as screening strain and turbidity as readout. Second, 16 confirmed hits were tested for their ability to induce the cell envelope stress responsive promoter piniBAC controlling expression of red fluorescent protein in an M. bovis BCG reporter strain. Using a fluorescence readout, the acetamide E11 was identified. Resistant mutant selection and whole genome sequencing revealed the mycolic acid transporter Mmpl3 as a candidate target of E11. Biochemical analysis using mycobacterial spheroplasts and various membrane assays suggest that E11 indirectly inhibits MmpL3-facilitated translocation of trehalose monomycolates by proton motive force disruption. E11 showed potent bactericidal activity against growing and non-growing M. tuberculosis, low cytotoxic, and hemolytic activity and a dynamic structure activity relationship. In addition to activity against M. tuberculosis, E11 was active against the non-tuberculous mycobacterium M. abscessus, an emerging opportunistic pathogen. In conclusion, we identified a novel bactericidal anti-mycobacterial lead compound targeting MmpL3 providing an attractive starting point for optimization. |
| first_indexed | 2024-12-11T04:21:41Z |
| format | Article |
| id | doaj.art-cb21df1d8aed41e0b481a951ca909fec |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T04:21:41Z |
| publishDate | 2018-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-cb21df1d8aed41e0b481a951ca909fec2022-12-22T01:21:05ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-12-01910.3389/fmicb.2018.02960424593Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force DisruptionAnnanya Shetty0Annanya Shetty1Zhujun Xu2Umayal Lakshmanan3Jeffrey Hill4Meng Ling Choong5Shu-Sin Chng6Shu-Sin Chng7Yoshiyuki Yamada8Anders Poulsen9Thomas Dick10Thomas Dick11Martin Gengenbacher12Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, SingaporeAntimicrobial Drug Discovery Laboratory, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, SingaporeDepartment of Chemistry, National University of Singapore, Singapore, SingaporeExperimental Therapeutics Center, A*STAR, Singapore, SingaporeExperimental Therapeutics Center, A*STAR, Singapore, SingaporeExperimental Therapeutics Center, A*STAR, Singapore, SingaporeDepartment of Chemistry, National University of Singapore, Singapore, SingaporeSingapore Center for Environmental Life Sciences Engineering, Nanyang Technological University, Singapore, SingaporeDepartment of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, SingaporeExperimental Therapeutics Center, A*STAR, Singapore, SingaporeAntimicrobial Drug Discovery Laboratory, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, SingaporePublic Health Research Institute, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ, United StatesPublic Health Research Institute, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ, United StatesTo identify novel inhibitors of Mycobacterium tuberculosis cell envelope biosynthesis, we employed a two-step approach. First, we screened the diverse synthetic small molecule 71,544-compound Enamine library for growth inhibitors using the non-pathogenic surrogate Mycobacterium bovis BCG as screening strain and turbidity as readout. Second, 16 confirmed hits were tested for their ability to induce the cell envelope stress responsive promoter piniBAC controlling expression of red fluorescent protein in an M. bovis BCG reporter strain. Using a fluorescence readout, the acetamide E11 was identified. Resistant mutant selection and whole genome sequencing revealed the mycolic acid transporter Mmpl3 as a candidate target of E11. Biochemical analysis using mycobacterial spheroplasts and various membrane assays suggest that E11 indirectly inhibits MmpL3-facilitated translocation of trehalose monomycolates by proton motive force disruption. E11 showed potent bactericidal activity against growing and non-growing M. tuberculosis, low cytotoxic, and hemolytic activity and a dynamic structure activity relationship. In addition to activity against M. tuberculosis, E11 was active against the non-tuberculous mycobacterium M. abscessus, an emerging opportunistic pathogen. In conclusion, we identified a novel bactericidal anti-mycobacterial lead compound targeting MmpL3 providing an attractive starting point for optimization.https://www.frontiersin.org/article/10.3389/fmicb.2018.02960/fullMycobacterium tuberculosisiniBACcell envelope stressflippasehigh throughput screen |
| spellingShingle | Annanya Shetty Annanya Shetty Zhujun Xu Umayal Lakshmanan Jeffrey Hill Meng Ling Choong Shu-Sin Chng Shu-Sin Chng Yoshiyuki Yamada Anders Poulsen Thomas Dick Thomas Dick Martin Gengenbacher Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption Frontiers in Microbiology Mycobacterium tuberculosis iniBAC cell envelope stress flippase high throughput screen |
| title | Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption |
| title_full | Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption |
| title_fullStr | Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption |
| title_full_unstemmed | Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption |
| title_short | Novel Acetamide Indirectly Targets Mycobacterial Transporter MmpL3 by Proton Motive Force Disruption |
| title_sort | novel acetamide indirectly targets mycobacterial transporter mmpl3 by proton motive force disruption |
| topic | Mycobacterium tuberculosis iniBAC cell envelope stress flippase high throughput screen |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.02960/full |
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