University of Oulu

Petäjä-Repo, U. E., Hogue, M., Laperrière, A., Walker, P., & Bouvier, M. (2000). Export from the Endoplasmic Reticulum Represents the Limiting Step in the Maturation and Cell Surface Expression of the Human δ Opioid Receptor. Journal of Biological Chemistry, 275(18), 13727–13736. https://doi.org/10.1074/jbc.275.18.13727

Export from the endoplasmic reticulum represents the limiting step in the maturation and cell surface expression of the human δ opioid receptor

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Author: Petäjä-Repo, Ulla E.1,2; Hogue, Mireille1; Laperrière, André1;
Organizations: 1Département de Biochimie, Université de Montréal, Montréal, Quebec H3C 3J7, Canada
2AstraZeneca R & D Montréal, St. Laurent, Quebec H4S 1Z9, Canada
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 0.3 MB)
Persistent link: http://urn.fi/urn:nbn:fi-fe202002115052
Language: English
Published: American Society for Biochemistry and Molecular Biology, 2000
Publish Date: 2020-02-11
Description:

Abstract

Synthesis and maturation of G protein-coupled receptors are complex events that require an intricate combination of processes that include protein folding, post-translational modifications, and transport through distinct cellular compartments. Relatively little is known about the nature and kinetics of specific steps involved in these processes. Here, the human δ opioid receptor expressed in human embryonic kidney 293S cells is used as a model to delineate these steps and to establish the kinetics of receptor synthesis, glycosylation, and transport. We found that the receptor is synthesized as a core-glycosylatedM r 45,000 precursor that is converted to the fully mature M r 55,000 receptor with a half-time of about 120 min. In addition to trimming and processing of two N-linked oligosaccharides, maturation involves addition of O-glycans containing N-acetylgalactosamine, galactose, and sialic acid. In contrast to N-glycosylation, which is initiated co-translationally and is completed when the protein reaches the trans-Golgi network, O-glycosylation was found to occur only after the receptor exits from the endoplasmic reticulum (ER) and was terminated as early as thetrans-Golgi cisternae. Once the carbohydrates are fully processed and the receptor reaches the trans-Golgi network, it is transported to the cell surface in about 10 min. The exit from the ER was found to be the limiting step in overall processing of the receptor. This indicates that early events in the folding of the receptor are probably rate-limiting and that receptor folding intermediates are retained in the ER until they can adopt the correct conformation. The overall low efficiency of receptor maturation, less than 50% of the precursor being processed to the fully glycosylated protein, further suggests that only a fraction of the synthesized receptors attain properly folded conformation that allows exit from the ER. This indicates that folding and ER export are key events in control of receptor cell surface expression. Whether or not the low efficiency of the ER export is a general feature among G protein-coupled receptors remains to be investigated.

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Series: Journal of biological chemistry
ISSN: 0021-9258
ISSN-E: 1083-351X
ISSN-L: 0021-9258
Volume: 275
Issue: 18
Pages: 13727 - 13736
DOI: 10.1074/jbc.275.18.13727
OADOI: https://oadoi.org/10.1074/jbc.275.18.13727
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Copyright information: This research was originally published in the Journal of Biological Chemistry. Petäjä-Repo, U. E., Hogue, M., Laperrière, A., Walker, P., & Bouvier, M. (2000). Export from the Endoplasmic Reticulum Represents the Limiting Step in the Maturation and Cell Surface Expression of the Human δ Opioid Receptor. J. Biol. Chem. 2000; 275:13737-13726. © the American Society for Biochemistry and Molecular Biology.