MmpL3 is a lipid transporter that binds trehalose monomycolate and phosphatidylethanolamine
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Abstract
The cell envelope of Mycobacterium tuberculosis is notable for the abundance of mycolic acids (MAs), essential to mycobacterial viability, and of other species-specific lipids. The mycobacterial cell envelope is extremely hydrophobic, which contributes to virulence and antibiotic resistance. However, exactly how fatty acids and lipidic elements are transported across the cell envelope for cell-wall biosynthesis is unclear. Mycobacterial membrane protein Large 3 (MmpL3) is essential and required for transport of trehalose monomycolates (TMMs), precursors of MA-containing trehalose dimycolates (TDM) and mycolyl arabinogalactan peptidoglycan, but the exact function of MmpL3 remains elusive. Here, we report a crystal structure of Mycobacterium smegmatis MmpL3 at a resolution of 2.59 Å, revealing a monomeric molecule that is structurally distinct from all known bacterial membrane proteins. A previously unknown MmpL3 ligand, phosphatidylethanolamine (PE), was discovered inside this transporter. We also show, via native mass spectrometry, that MmpL3 specifically binds both TMM and PE, but not TDM, in the micromolar range. These observations provide insight into the function of MmpL3 and suggest a possible role for this protein in shuttling a variety of lipids to strengthen the mycobacterial cell wall.
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Inhibitors block the proton translocation channel of MmpL3 to arrest TMM translocation.
“how inhibitors block this proton translocation channel to arrest TMM translocation”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [155]
The Proposed Mechanism for TMM Translocation section describes TMM moving through an MmpL3 channel, energized by a proton-relay network. The MmpL3 Specifically Binds a Variety of Lipids section says potential therapeutics can block lipid binding and transport, but does not say inhibitors block the proton-translocation pathway specifically. The claim's mechanism of inhibition is therefore unsupported. (LLM, conf=0.84)
Experimental — accuracy not yet measuredX-ray crystallography structures of M. smegmatis MmpL3 alone and complexed with its ligand TMM and known inhibitors were solved.
“To gain a molecular view of the MmpL3 floppase and the mechanism of drug inhibition, X-ray crystallography structures of M. smegmatis MmpL3 alone and complexed with its ligand TMM and known inhibitors were solved (154, 155).”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [155]
The Results report X-ray crystal structures of M. smegmatis MmpL3, but identify PE and DDM—not TMM—as bound in the structure (page unavailable). TMM binding was shown by native mass spectrometry. The paper credits Zhang et al. with structures involving several presumed inhibitors; it does not report solving those inhibitor-bound structures itself. (LLM, conf=0.91)
Experimental — accuracy not yet measuredMmpL3 utilizes a proton gradient to drive TMM translocation.
“These structures revealed the mechanism by which the protein utilizes a proton gradient to drive TMM translocation and how inhibitors block this proton translocation channel to arrest TMM translocation.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [155]
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Mycobacterium smegmatis: The Vanguard of Mycobacterial Research
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- Authors
- Su, C.-C., Klenotic, P. A., Bolla, J. R., Purdy, G. E., Robinson, C. V., & Yu, E. W.
- Affiliations
- Case Western Reserve University (US)
- University of Oxford (GB)
- Oregon Health & Science University (US)
- ORCID
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2019-05-21
- Volume
- 116
- Issue
- 23
- Pages
- 11241-11246
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC6561238
- OpenAlex
- W2913841228
- Cited by
- 148
- References
- 45
- Citations by year
- 2026: 12 · 2025: 20 · 2024: 24 · 2023: 22 · 2022: 13 · 2021: 25 · 2020: 23 · 2019: 9
- FWCI
- 13.35
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- Office of Extramural Research, National Institutes of Health (AI123148, AI145069)
- RCUK | Medical Research Council (MR/N020413/1)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Keywords
- MmpL3, Mycobacterium tuberculosis, Mycobacterium smegmatis, lipid transfer proteins, trehalose monomycolate, phosphatidylethanolamine, mycolic acids, trehalose dimycolate, cell envelope, membrane protein crystal structure, lipid transport, cell wall biosynthesis
- MeSH terms
- Bacterial Proteins, Biological Transport, Cell Membrane, Cell Wall, Cord Factors, Mycolic Acids, Phosphatidylethanolamines, Mycobacterium smegmatis, Membrane Transport Proteins
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