Functional expression of opioid receptors and other human GPCRs in yeast engineered to produce human sterols.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
24 05 2022
24 05 2022
Historique:
received:
14
12
2021
accepted:
09
05
2022
entrez:
24
5
2022
pubmed:
25
5
2022
medline:
27
5
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 35610225
doi: 10.1038/s41467-022-30570-7
pii: 10.1038/s41467-022-30570-7
pmc: PMC9130329
doi:
Substances chimiques
Phytosterols
0
Receptors, Opioid
0
Receptors, Opioid, kappa
0
Receptors, Opioid, mu
0
Sterols
0
Cholesterol
97C5T2UQ7J
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
2882Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM122480
Pays : United States
Organisme : NCCIH NIH HHS
ID : R21 AT010777
Pays : United States
Informations de copyright
© 2022. The Author(s).
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