Mapping of Mycobacterial Enzymes Involved in Triacylglycerol Accumulation as Intrabacterial Lipid Inclusions Using Activity-Based Multitarget Inhibitor Probes
Lipids play a critical role in the physiology, life cycle, and pathogenicity of mycobacteria. They largely participate in host-pathogen interactions and fulfill important functions ranging from cell wall biosynthesis/maintenance, bacterial growth dynamics, and long-term persistence. In that context, triacylglycerol, a specific subtype of neutral lipid, is stored as intrabacterial lipid inclusions (ILI), which have been described as important structures for long-term survival and persistence within the host. However, the enzymes involved in ILI formation and degradation remain largely undefined. Using bio-orthogonal click-chemistry activity-based protein profiling (CC-ABPP) of newly synthesized oxadiazolone (<b>OX</b>), cyclophostin, and cyclipostins (<b>CyC</b>) probes, we report the direct capture of target proteins in <i>Mycobacterium abscessus</i> growing under carbon excess and nitrogen-deprived in vitro conditions that promote triacylglycerol production and ILI formation. This approach led to the identification of a set of 65 enzymes potentially involved in global processes related to ILI anabolism. Among these, the long-chain-fatty-acid--CoA ligase MAB_1978c/FadD15 has been validated as a pivotal enzyme that colocalizes on ILI and as a major contributor in ILI formation in <i>M. abscessus</i>.
Authors
- Stéphane Audebert (ORCID: https://orcid.org/0000-0002-9409-2588)
- Tonia Dargham
- Ladine Çelik (ORCID: https://orcid.org/0000-0003-3352-9181)
- Luc Camoin (ORCID: https://orcid.org/0000-0002-1230-4787)
- Thierry Durand (ORCID: https://orcid.org/0000-0001-6086-7296)
- Romain Avellan
- Céline Crauste (ORCID: https://orcid.org/0000-0002-5714-8749)
- Laurent Kremer (ORCID: https://orcid.org/0000-0002-6604-4458)
- Jordan Lehoux (ORCID: https://orcid.org/0000-0001-7369-0266)
- Stéphane Canaan (ORCID: https://orcid.org/0000-0001-7478-300X)
- Pierre Santucci (ORCID: https://orcid.org/0000-0002-6291-3425)
- Vanessa Point
- Christopher D. Spilling
- Isabelle Poncin
- Jean‐François Cavalier (ORCID: https://orcid.org/0000-0003-0864-8314)
- Alexandre Guy (ORCID: https://orcid.org/0000-0001-8206-8946)
- Thomas Francis
Institutions
- École Nationale Supérieure de Chimie de Montpellier (FR)
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Université de Montpellier (FR)
- University of Missouri–St. Louis (US)
- Aix-Marseille Université (FR)
- Méditerranée Infection Foundation (FR)
- Institut de Recherche en Infectiologie de Montpellier (FR)
- Institut des Biomolécules Max Mousseron (FR)
- Institut Paoli-Calmettes (FR)
Publication Details
- Journal
- ACS Infectious Diseases
- Published
- 2025-05-29
- DOI
- https://doi.org/10.1021/acsinfecdis.5c00127
- Primary Topic
- Mycobacterium research and diagnosis
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Fondation Méditerranée Infection
- Aix-Marseille Université
- Agence Nationale de la Recherche
- Ministère de l'Education Nationale, de l'Enseignement Superieur et de la Recherche
- Centre National de la Recherche Scientifique