Functional expression of opioid receptors and other human GPCRs in yeast engineered to produce human sterols

Björn D. M. Bean, Colleen J. Mulvihill, Riddhiman K. Garge, Daniel R. Boutz, Olivier Rousseau, Brendan M. Floyd, William Cheney, Elizabeth C. Gardner, Andrew D. Ellington, Edward M. Marcotte, Jimmy D. Gollihar (), Malcolm Whiteway and Vincent J. J. Martin ()
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Björn D. M. Bean: Concordia University
Colleen J. Mulvihill: The University of Texas at Austin
Riddhiman K. Garge: The University of Texas at Austin
Daniel R. Boutz: The University of Texas at Austin
Olivier Rousseau: Concordia University
Brendan M. Floyd: The University of Texas at Austin
William Cheney: Concordia University
Elizabeth C. Gardner: The University of Texas at Austin
Andrew D. Ellington: The University of Texas at Austin
Edward M. Marcotte: The University of Texas at Austin
Jimmy D. Gollihar: The University of Texas at Austin
Malcolm Whiteway: Concordia University
Vincent J. J. Martin: Concordia University

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: 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.

Date: 2022
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-30570-7

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DOI: 10.1038/s41467-022-30570-7

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