Site-specific O-glycosylation of N-terminal serine residues by polypeptide GalNAc-transferase 2 modulates human δ-opioid receptor turnover at the plasma membrane

Cellular Signalling
Jarkko J LackmanUlla E Petäjä-Repo

Abstract

G protein-coupled receptors (GPCRs) are an important protein family of signalling receptors that govern a wide variety of physiological functions. The capacity to transmit extracellular signals and the extent of cellular response are largely determined by the amount of functional receptors at the cell surface that is subject to complex and fine-tuned regulation. Here, we demonstrate that the cell surface expression level of an inhibitory GPCR, the human δ-opioid receptor (hδOR) involved in pain and mood regulation, is modulated by site-specific N-acetylgalactosamine (GalNAc) -type O-glycosylation. Importantly, we identified one out of the 20 polypeptide GalNAc-transferase isoforms, GalNAc-T2, as the specific regulator of O-glycosylation of Ser6, Ser25 and Ser29 in the N-terminal ectodomain of the receptor. This was demonstrated by in vitro glycosylation assays using peptides corresponding to the hδOR N-terminus, Vicia villosa lectin affinity purification of receptors expressed in HEK293 SimpleCells capable of synthesizing only truncated O-glycans, GalNAc-T edited cell line model systems, and site-directed mutagenesis of the putative O-glycosylation sites. Interestingly, a single-nucleotide polymorphism, at residue 27 (F27C), wa...Continue Reading

Citations

Jan 17, 2019·Journal of Biochemistry·Naosuke Nakamura, Akira Kurosaka
Mar 30, 2019·Journal of Peptide Science : an Official Publication of the European Peptide Society·Claudia BelloAnna Maria Papini
Apr 6, 2019·The Journal of Biological Chemistry·David SalomKrzysztof Palczewski
Nov 22, 2020·Biomolecules·Keiko Akasaka-Manya, Hiroshi Manya
Feb 23, 2021·The Journal of Biological Chemistry·Yoshiki NarimatsuHenrik Clausen
May 28, 2020·Trends in Neurosciences·Mariana Lemos Duarte, Lakshmi A Devi
Apr 17, 2020·ACS Pharmacology & Translational Science·Christoffer K GothMette M Rosenkilde
Jun 24, 2020·Peptides·Richard J Bodnar
Mar 9, 2019·Langmuir : the ACS Journal of Surfaces and Colloids·Yuzhen WangYan Fang
Sep 25, 2021·Journal of Neuroscience Research·Jade DegrandmaisonLouis Gendron

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