Biochemical and Biological Assays of Mycolactone-Mediated Inhibition of Sec61

Sarah O'Keefe, Caroline Demangel, Stephen High

Tutkimustuotos: Artikkeli kirjassa/raportissa/konferenssijulkaisussaKirjan luku tai artikkeliTieteellinenvertaisarvioitu

Abstrakti

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.

Alkuperäiskielienglanti
OtsikkoMethods in Molecular Biology
ToimittajatGerd Pluschke, Katharina Röltgen
Sivumäärä19
KustantajaHumana
Julkaisupäivä14 lokak. 2021
Sivut163-181
ISBN (painettu)978-1-0716-1778-6
ISBN (elektroninen)978-1-0716-1779-3
DOI - pysyväislinkit
TilaJulkaistu - 14 lokak. 2021
Julkaistu ulkoisestiKyllä
OKM-julkaisutyyppiA3 Kirjan tai muun kokoomateoksen osa

Julkaisusarja

NimiMethods in Molecular Biology
Vuosikerta2387
ISSN (painettu)1064-3745
ISSN (elektroninen)1940-6029

Lisätietoja

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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