Direct Inhibition of MmpL3 by Novel Antitubercular Compounds
Open accessAbstract
MmpL3, an essential transporter involved in the export of mycolic acids, is the proposed target of a number of antimycobacterial inhibitors under development. Whether MmpL3 serves as the direct target of these compounds, however, has been called into question after the discovery that some of them dissipated the proton motive force from which MmpL transporters derive their energy. Using a combination of in vitro and whole-cell-based approaches, we here provide evidence that five structurally distinct MmpL3 inhibitor series, three of which impact proton motive force in Mycobacterium tuberculosis, directly interact with MmpL3. Medium- to high-throughput assays based on these approaches were developed to facilitate the future screening and optimization of MmpL3 inhibitors. The promiscuity of MmpL3 as a drug target and the mechanisms through which missense mutations located in a transmembrane region of this transporter may confer cross-resistance to a variety of chemical scaffolds are discussed in light of the exquisite vulnerability of MmpL3, its apparent mechanisms of interaction with inhibitors, and evidence of conformational changes induced both by the inhibitors and one of the most commonly identified resistance mutations in MmpL3.
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The M. smegmatis spheroplast assay established the first specific platform for validating other potential MmpL3 inhibitors.
“While this study established the first, specific platform for validating other potential MmpL3 inhibitors, the technical difficulty involved with creating spheroplasts prompted development of a more high-throughput approach (153).”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
The Abstract (page not provided) says the M. smegmatis spheroplast assay gave the first direct evidence of TMM buildup following MmpL3 inhibition, not a specific platform for validating potential inhibitors. It explicitly says the assay cannot readily distinguish direct from indirect inhibition; it does support the citing sentence’s point that spheroplast preparation is cumbersome. (LLM, conf=0.88)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredUndiscovered MmpL3 inhibitors would most likely bind the same common active site.
“predicting that undiscovered inhibitors would most likely bind this same site”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
The Abstract concludes that the inhibitors studied share a common binding site, and the Discussion describes three known inhibitor series binding at the same site. Neither passage predicts that undiscovered inhibitors would most likely bind there; that extension goes beyond the paper's stated findings. Page numbers were not available in our copy. (LLM, conf=0.83)
Experimental — accuracy not yet measuredThe mmpL3 mutations conferred cross-resistance to different drugs.
“Unexpectedly, the mmpL3 mutations conferred cross-resistance to different drugs, suggesting that most inhibitors bind a common active site, and predicting that undiscovered inhibitors would most likely bind this same site.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
M. smegmatis was used as a surrogate to express different drug-resistant M. tuberculosis mmpL3 alleles.
“M. smegmatis was used as a surrogate to express different drug-resistant M. tuberculosis mmpL3 alleles.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
Technical difficulty in creating spheroplasts prompted development of a more high-throughput approach.
“the technical difficulty involved with creating spheroplasts prompted development of a more high-throughput approach (153)”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
Most MmpL3 inhibitors bind a common active site.
“suggesting that most inhibitors bind a common active site”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [153]
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Mycobacterium smegmatis: The Vanguard of Mycobacterial Research
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- Authors
- Li, W., Stevens, C. M., Pandya, A. N., Darzynkiewicz, Z., Bhattarai, P., Tong, W., Gonzalez-Juarrero, M., North, E. J., Zgurskaya, H. I., & Jackson, M.
- Affiliations
- Colorado State University (US)
- University of Oklahoma (US)
- Creighton University (US)
- University of Oklahoma , , 101 Stephenson Parkway , , ,
- Colorado State University , , 1682 Campus Delivery , , ,
- Corresponding author
- Zgurskaya, Helen I.
- Jackson, Mary
- ORCID
- Venue
- ACS Infectious Diseases
- Publisher
- American Chemical Society (ACS)
- Published
- 2019-03-18
- Volume
- 5
- Issue
- 6
- Pages
- 1001-1012
- ISSN
- 2373-8227 (ISSN-L)
- PMC
- PMC6580365
- OpenAlex
- W2921257501
- Cited by
- 97
- References
- 34
- Citations by year
- 2026: 7 · 2025: 14 · 2024: 10 · 2023: 15 · 2022: 13 · 2021: 9 · 2020: 20 · 2019: 9
- FWCI
- 9.77
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- ACS Infectious Diseases (publishedVersion, cc-by)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (AI116525)
- Bill and Melinda Gates Foundation (OPP1181207)
- Global Alliance for TB Drug Development (NA)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Antibiotic Resistance in Bacteria
- Keywords
- MmpL3, Mycobacterium tuberculosis, antitubercular agents, proton motive force, transporter inhibition, drug target validation, missense mutations, cross-resistance, conformational change, high-throughput assay
- MeSH terms
- Drug Development, Antitubercular Agents, Bacterial Proteins, Humans, Microbial Sensitivity Tests, Mycobacterium tuberculosis, Proton-Motive Force, Membrane Transport Proteins
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