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, Vincent J J Martin

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article number2882
Pages (from-to)2882
JournalNature Communications
Volume13
Issue number1
DOIs
StatePublished - May 24 2022

Keywords

  • Cholesterol/metabolism
  • Humans
  • Phytosterols/metabolism
  • Receptors, Opioid/metabolism
  • Receptors, Opioid, kappa/agonists
  • Receptors, Opioid, mu/metabolism
  • Saccharomyces cerevisiae/genetics
  • Sterols/metabolism

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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