Photoactivatable Glycolipid Probes for Identifying Mycolate-Protein Interactions in Live Mycobacteria.

TitlePhotoactivatable Glycolipid Probes for Identifying Mycolate-Protein Interactions in Live Mycobacteria.
Publication TypeJournal Article
Year of Publication2020
AuthorsKavunja HW, Biegas KJ, Banahene N, Stewart JA, Piligian BF, Groenevelt JM, Sein CE, Morita YS, Niederweis M, M Siegrist S, Swarts BM
JournalJ Am Chem Soc
Date Published2020 04 29

Mycobacteria have a distinctive glycolipid-rich outer membrane, the mycomembrane, which is a critical target for tuberculosis drug development. However, proteins that associate with the mycomembrane, or that are involved in its metabolism and host interactions, are not well-characterized. To facilitate the study of mycomembrane-related proteins, we developed photoactivatable trehalose monomycolate analogues that metabolically incorporate into the mycomembrane in live mycobacteria, enabling photo-cross-linking and click-chemistry-mediated analysis of mycolate-interacting proteins. When deployed in with quantitative proteomics, this strategy enriched over 100 proteins, including the mycomembrane porin (MspA), several proteins with known mycomembrane synthesis or remodeling functions (CmrA, MmpL3, Ag85, Tdmh), and numerous candidate mycolate-interacting proteins. Our approach is highly versatile, as it (i) enlists click chemistry for flexible protein functionalization; (ii) in principle can be applied to any mycobacterial species to identify endogenous bacterial proteins or host proteins that interact with mycolates; and (iii) can potentially be expanded to investigate protein interactions with other mycobacterial lipids. This tool is expected to help elucidate fundamental physiological and pathological processes related to the mycomembrane and may reveal novel diagnostic and therapeutic targets.

Alternate JournalJ Am Chem Soc
PubMed ID32293873
Grant ListDP2 AI138238 / AI / NIAID NIH HHS / United States
R21 AI144748 / AI / NIAID NIH HHS / United States
R03 AI140259 / AI / NIAID NIH HHS / United States
R01 AI121354 / AI / NIAID NIH HHS / United States