N-glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants |
|
Author: | Lackman, Jarkko J.1; Markkanen, Piia M. H.1; Hogue, Mireille2; |
Organizations: |
1Department of Anatomy and Cell Biology and the Medical Research Center Oulu, 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 |
Format: | article |
Version: | published version |
Access: | open |
Online Access: | PDF Full Text (PDF, 1.7 MB) |
Persistent link: | http://urn.fi/urn:nbn:fi-fe2019120545771 |
Language: | English |
Published: |
American Society for Biochemistry and Molecular Biology,
2014
|
Publish Date: | 2019-12-05 |
Description: |
AbstractQuality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys²⁷ to ERAD. However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys²⁷ reached the cell surface in conformation incompatible with ligand binding. The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys²⁷, pointing to an important role of CNX in the hδOR N-glycan-dependent QC. This was further supported by the fact that inhibiting the co-translational interaction of hδOR-Cys²⁷ precursors with CNX led to their ERAD. Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys²⁷ variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. see all
|
Series: |
Journal of biological chemistry |
ISSN: | 0021-9258 |
ISSN-E: | 1083-351X |
ISSN-L: | 0021-9258 |
Volume: | 289 |
Issue: | 25 |
Pages: | 17830 - 17842 |
DOI: | 10.1074/jbc.M114.566273 |
OADOI: | https://oadoi.org/10.1074/jbc.M114.566273 |
Type of Publication: |
A1 Journal article – refereed |
Field of Science: |
1182 Biochemistry, cell and molecular biology |
Subjects: | |
Funding: |
This work was supported by the Sigrid Jusélius Foundation, the Medical Research Center Oulu and Grant 127199 from the Academy of Finland (to U. E. P.-R.). Jarkko J. Lackman was supported by the Finnish Cultural Foundation. |
Academy of Finland Grant Number: |
127199 |
Detailed Information: |
127199 (Academy of Finland Funding decision) |
Copyright information: |
This research was originally published in the Journal of Biological Chemistry. Lackman, J. J., Markkanen, P. M. H., Hogue, M., Bouvier, M., & Petäjä-Repo, U. E.. N-Glycan-dependent and -independent Quality Control of Human δ Opioid Receptor N-terminal Variants. J. Biol. Chem. 2012; 289:17830-17842. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. |