University of Oulu

Leskelä, T. T., Markkanen, P. M. H., Pietilä, E. M., Tuusa, J. T., & Petäjä-Repo, U. E. (2007). Opioid Receptor Pharmacological Chaperones Act by Binding and Stabilizing Newly Synthesized Receptors in the Endoplasmic Reticulum. Journal of Biological Chemistry, 282(32), 23171–23183. https://doi.org/10.1074/jbc.m610896200

Opioid receptor pharmacological chaperones act by binding and stabilizing newly synthesized receptors in the endoplasmic reticulum

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Author: Leskelä, Tarja T.1; Markkanen, Piia M. H.1; Pietilä, E. Maritta1;
Organizations: 1Biocenter Oulu and Department of Anatomy and Cell Biology, University of Oulu, FI-90014 Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 0.8 MB)
Persistent link: http://urn.fi/urn:nbn:fi-fe2019121948927
Language: English
Published: American Society for Biochemistry and Molecular Biology, 2007
Publish Date: 2019-12-19
Description:

Abstract

Accumulating evidence has indicated that membrane-permeable G protein-coupled receptor ligands can enhance cell surface targeting of their cognate wild-type and mutant receptors. This pharmacological chaperoning was thought to result from ligand-mediated stabilization of immature receptors in the endoplasmic reticulum (ER). In the present study, we directly tested this hypothesis using wild-type and mutant forms of the human δ-opioid receptor as models. ER-localized receptors were isolated by expressing the receptors in HEK293 cells under tightly controlled tetracycline induction and blocking their ER export with brefeldin A. The ER-retained δ-opioid receptor precursors were able to bind [³H]diprenorphine with high affinity, and treatment of cells with an opioid antagonist naltrexone led to a 2-fold increase in the number of binding sites. After removing the transport block, the antagonist-mediated increase in the number of receptors was detectable at the cell surface by flow cytometry and cell surface biotinylation assay. Importantly, opioid ligands, both antagonists and agonists, were found to stabilize the ER-retained receptor precursors in an in vitro heat inactivation assay and the treatment enhanced dissociation of receptor precursors from the molecular chaperone calnexin. Thus, we conclude that pharmacological chaperones facilitate plasma membrane targeting of δ-opioid receptors by binding and stabilizing receptor precursors, thereby promoting their release from the stringent ER quality control.

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Series: Journal of biological chemistry
ISSN: 0021-9258
ISSN-E: 1083-351X
ISSN-L: 0021-9258
Volume: 282
Issue: 32
Pages: 23171 - 23183
DOI: 10.1074/jbc.M610896200
OADOI: https://oadoi.org/10.1074/jbc.M610896200
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Funding: This work was supported by the Biocenter Oulu and by Academy of Finland Grant 200732. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Academy of Finland Grant Number: 200732
Detailed Information: 200732 (Academy of Finland Funding decision)
Copyright information: This research was originally published in the Journal of Biological Chemistry. Leskelä, T. T., Markkanen, P. M. H., Pietilä, E. M., Tuusa, J. T., & Petäjä-Repo, U. E.. Opioid Receptor Pharmacological Chaperones Act by Binding and Stabilizing Newly Synthesized Receptors in the Endoplasmic Reticulum. J. Biol. Chem. Year; 282:23171-23183. © the American Society for Biochemistry and Molecular Biology.