Leskelä, T. T., Lackman, J. J., Vierimaa, M. M., Kobayashi, H., Bouvier, M., & Petäjä-Repo, U. E. (2011). Cys-27 Variant of Human δ-Opioid Receptor Modulates Maturation and Cell Surface Delivery of Phe-27 Variant via Heteromerization. Journal of Biological Chemistry, 287(7), 5008–5020. https://doi.org/10.1074/jbc.m111.305656
Cys-27 variant of human delta-opioid receptor modulates maturation and cell surface delivery of Phe-27 variant via heteromerization
|Author:||Leskelä, Tarja T.1; Lackman, Jarkko J.1; Vierimaa, Miia M.1;|
1Department of Anatomy and Cell Biology, Institute of Biomedicine, University of Oulu, FI-90014 Oulu, Finland
2Department of Biochemistry, Institute for Research in Immunology and Cancer, and Groupe de Recherche Universitaire sur le Médicament, Université de Montréal, Montréal, Québec H3C 3J7, Canada
|Online Access:||PDF Full Text (PDF, 1.6 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2019120445647
American Society for Biochemistry and Molecular Biology,
|Publish Date:|| 2019-12-04
The important role of G protein-coupled receptor homo/heteromerization in receptor folding, maturation, trafficking, and cell surface expression has become increasingly evident. Here we investigated whether the human δ-opioid receptor (hδOR) Cys-27 variant that shows inherent compromised maturation has an effect on the behavior of the more common Phe-27 variant in the early secretory pathway. We demonstrate that hδOR-Cys-27 acts in a dominant negative manner and impairs cell surface delivery of the co-expressed hδOR-Phe-27 and impairs conversion of precursors to the mature form. This was demonstrated by metabolic labeling, Western blotting, flow cytometry, and confocal microscopy in HEK293 and human SH-SY5Y neuroblastoma cells using differentially epitope-tagged variants. The hδOR-Phe-27 precursors that were redirected to the endoplasmic reticulum-associated degradation were, however, rescued by a pharmacological chaperone, the opioid antagonist naltrexone. Co-immunoprecipitation of metabolically labeled variants revealed that both endoplasmic reticulum-localized precursors and mature receptors exist as homo/heteromers. The existence of homo/heteromers was confirmed in living cells by bioluminescence resonance energy transfer measurements, showing that the variants have a similar propensity to form homo/heteromers. By forming both homomers and heteromers, the hδOR-Cys-27 variant may thus regulate the levels of receptors at the cell surface, possibly leading to altered responsiveness to opioid ligands in individuals carrying the Cys-27 variant.
Journal of biological chemistry
|Pages:||5008 - 5020|
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
This work was supported by the Sigrid Jusélius Foundation and Academy of Finland Grant 127199 (to U. E. P.-R.) and Canadian Institutes for Health Research Grant MOP-11215 (to M. B.). Tarja T. Leskelä was supported by the Finnish Cultural Foundation. Jarkko J. Lackman was supported by the Finnish Glycoscience Graduate School.
|Academy of Finland Grant Number:||
127199 (Academy of Finland Funding decision)
This research was originally published in the Journal of Biological Chemistry. Leskelä, T. T., Lackman, J. J., Vierimaa, M. M., Kobayashi, H., Bouvier, M., & Petäjä-Repo, U. E.. Cys-27 Variant of Human δ-Opioid Receptor Modulates Maturation and Cell Surface Delivery of Phe-27 Variant via Heteromerization. J. Biol. Chem. 2012; 287:5008-5020. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.