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© Institut Pasteur/Antoinette Ryter
Coupe de Mycobacterium bovis ou bacille de Calmette et Guérin (BCG). Souche atténuée de bacille vivant, à l'origine du vaccin antituberculeux délivré par voie intradermique ou scarifications (Grossissement X 70000). Image colorisée.
Publication : Methods in molecular biology (Clifton, N.J.)

Biochemical and Biological Assays of Mycolactone-Mediated Inhibition of Sec61.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2022

O'Keefe S, High S, Demangel C,

Link to Pubmed [PMID] – 34643911

Link to DOI – 10.1007/978-1-0716-1779-3_16

Methods Mol Biol 2022 ; 2387(): 163-181

Mycobacterium ulcerans, the causative agent of Buruli ulcer disease, is unique among human pathogens in its capacity to produce mycolactone, a diffusible macrolide with immunosuppressive and cytotoxic properties. Recent studies have shown that mycolactone operates by inhibiting the host membrane translocation complex (Sec61), with an unprecedented potency compared to previously identified Sec61 blockers. Mycolactone binding to the pore-forming subunit of Sec61 inhibits its capacity to transport nascent secretory and membrane proteins into the endoplasmic reticulum, leading to their cytosolic degradation by the ubiquitin:proteasome system. In T lymphocytes, Sec61 blockade by mycolactone manifests as a sharp decrease in the cell’s ability to express homing receptors and release cytokines following activation. Sustained exposure of human cells to mycolactone typically generates proteotoxic stress responses in their cytosol and endoplasmic reticulum (ER), ultimately inducing apoptosis. Here we describe cell-free systems for studying Sec61-mediated protein translocation that allow the impact of mycolactone on the biogenesis of secretory and membrane proteins to be probed. We also describe biological assays of mycolactone-driven inhibition of Sec61 providing rapid and sensitive means to quantitatively assess the presence of the toxin in biological samples.

https://pubmed.ncbi.nlm.nih.gov/34643911