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- Title
Functional expression of opioid receptors and other human GPCRs in yeast engineered to produce human sterols.
- Authors
Bean, Björn D. M.; Mulvihill, Colleen J.; Garge, Riddhiman K.; Boutz, Daniel R.; Rousseau, Olivier; Floyd, Brendan M.; Cheney, William; Gardner, Elizabeth C.; Ellington, Andrew D.; Marcotte, Edward M.; Gollihar, Jimmy D.; Whiteway, Malcolm; Martin, Vincent J. J.
- Abstract
The yeast Saccharomyces cerevisiae is powerful for studying human G protein-coupled receptors as they can be coupled to its mating pathway. However, some receptors, including the mu opioid receptor, are non-functional, which may be due to the presence of the fungal sterol ergosterol instead of cholesterol. Here we engineer yeast to produce cholesterol and introduce diverse mu, delta, and kappa opioid receptors to create sensitive opioid biosensors that recapitulate agonist binding profiles and antagonist inhibition. Additionally, human mu opioid receptor variants, including those with clinical relevance, largely display expected phenotypes. By testing mu opioid receptor-based biosensors with systematically adjusted cholesterol biosynthetic intermediates, we relate sterol profiles to biosensor sensitivity. Finally, we apply sterol-modified backgrounds to other human receptors revealing sterol influence in SSTR5, 5-HTR4, FPR1, and NPY1R signaling. This work provides a platform for generating human G protein-coupled receptor-based biosensors, facilitating receptor deorphanization and high-throughput screening of receptors and effectors. The yeast Saccharomyces cerevisiae is powerful for studying human G protein-coupled receptors as they can be coupled to its mating pathway. Here the authors engineer baker's yeast to produce human sterols and show that vertebrate G protein coupled receptors are more sensitive in this membrane environment.
- Subjects
OPIOID receptors; G protein coupled receptors; STEROLS; ERGONOMICS; YEAST; SACCHAROMYCES cerevisiae
- Publication
Nature Communications, 2022, Vol 13, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-022-30570-7