A Screen for Protein–Protein Interactions in Live Mycobacteria Reveals a Functional Link between the Virulence-Associated Lipid Transporter LprG and the Mycolyltransferase Antigen 85A
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Outer membrane lipids in pathogenic mycobacteria are important for virulence and survival. Although the biosynthesis of these lipids has been extensively studied, mechanisms responsible for their assembly in the outer membrane are not understood. In the study of Gram-negative outer membrane assembly, protein-protein interactions define transport mechanisms, but analogous interactions have not been explored in mycobacteria. Here we identified interactions with the lipid transport protein LprG. Using site-specific photo-cross-linking in live mycobacteria, we mapped three major interaction interfaces within LprG. We went on to identify proteins that cross-link at the entrance to the lipid binding pocket, an area likely relevant to LprG transport function. We verified LprG site-specific interactions with two hits, the conserved lipoproteins LppK and LppI. We further showed that LprG interacts physically and functionally with the mycolyltransferase Ag85A, as loss of either protein leads to similar defects in cell growth and mycolylation. Overall, our results support a model in which protein interactions coordinate multiple pathways in outer membrane biogenesis and connect lipid biosynthesis to transport.
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Identifying protein-protein interactions can provide insight on protein function.
“Identifying protein-protein interactions can provide enormous insight on protein function, as demonstrated by new in vivo proximity-labeling technologies.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [160]
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- [44]Nguyen L Chinnapapagari S Thompson CJ FbpA-Dependent biosynthesis of trehalose dimycolate is required for the intrinsic multidrug resistance, cell wall structure, and colonial morphology of Mycobacterium smegmatis J Bacteriol 2005 187 19 6603 11 10.1128/JB.187.19.6603-6611.2005 16166521 PMC1251576 · DOI · Paper page
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- Authors
- Touchette, M. H., Van Vlack, E. R., Bai, L., Kim, J., Cognetta, A. B., Previti, M. L., Backus, K. M., Martin, D. W., Cravatt, B. F., & Seeliger, J. C.
- Affiliations
- Stony Brook University (US)
- Department of Chemistry, Stony Brook University, Stony Brook, New York 11790, United States
- Scripps Research Institute (US)
- Corresponding author
- Seeliger, Jessica C.
- ORCID
- Venue
- ACS Infectious Diseases
- Publisher
- American Chemical Society (ACS)
- Published
- 2017-03-21
- Volume
- 3
- Issue
- 5
- Pages
- 336-348
- ISSN
- 2373-8227 (ISSN-L)
- PMC
- PMC5632815
- OpenAlex
- W2594126074
- Cited by
- 34
- References
- 49
- Citations by year
- 2026: 3 · 2025: 5 · 2024: 2 · 2023: 5 · 2022: 1 · 2021: 6 · 2020: 5 · 2019: 4 · 2018: 3
- FWCI
- 2.53
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- ACS Infectious Diseases (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Center for Research Resources (1 S10 RR023680-1)
- National Institute of General Medical Sciences (F32 GM108208)
- Stony Wold-Herbert Fund
- State University of New York
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Keywords
- Mycobacterium tuberculosis, protein-protein interaction, Ag85A, mycolic acid biosynthesis, lipid transport, outer membrane biogenesis, virulence, live-cell screening
- MeSH terms
- Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Acyltransferases, Antigens, Bacterial, Bacterial Proteins, Binding Sites, Carrier Proteins, Cell Membrane, Cloning, Molecular, Kinetics, Lipids, Models, Molecular, Mycobacterium tuberculosis, Plasmids, Protein Binding, Recombinant Proteins, Virulence, Gene Expression, Mycobacterium smegmatis, Amino Acid Motifs, Protein Interaction Domains and Motifs
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