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タイトル: | Identification and functional characterisation of N-linked glycosylation of the orphan G protein-coupled receptor Gpr176 |
著者: | Wang, Tianyu Nakagawa, Shumpei Miyake, Takahito https://orcid.org/0000-0002-4356-5883 (unconfirmed) Setsu, Genzui Kunisue, Sumihiro Goto, Kaoru Hirasawa, Akira https://orcid.org/0000-0001-5692-805X (unconfirmed) Okamura, Hitoshi Yamaguchi, Yoshiaki https://orcid.org/0000-0003-4126-542X (unconfirmed) Doi, Masao https://orcid.org/0000-0001-6264-9217 (unconfirmed) |
著者名の別形: | 三宅, 崇仁 平澤, 明 岡村, 均 山口, 賀章 土居, 雅夫 |
キーワード: | Circadian rhythms and sleep Glycosylation Hormone receptors Mutation Post-translational modifications |
発行日: | 10-Mar-2020 |
出版者: | Springer Nature |
誌名: | Scientific Reports |
巻: | 10 |
論文番号: | 4429 |
抄録: | G-protein-coupled receptors (GPCRs) are important drug targets with diverse therapeutic applications. However, there are still more than a hundred orphan GPCRs, whose protein functions and biochemical features remain unidentified. Gpr176 encodes a class-A orphan GPCR that has a role in circadian clock regulation in mouse hypothalamus and is also implicated in human breast cancer transcriptional response. Here we show that Gpr176 is N-glycosylated. Peptide-N-glycosidase treatment of mouse hypothalamus extracts revealed that endogenous Gpr176 undergoes N-glycosylation. Using a heterologous expression system, we show that N-glycosylation occurs at four conserved asparagine residues in the N-terminal region of Gpr176. Deficient N-glycosylation due to mutation of these residues reduced the protein expression of Gpr176. At the molecular function level, Gpr176 has constitutive, agonist-independent activity that leads to reduced cAMP synthesis. Although deficient N-glycosylation did not compromise this intrinsic activity, the resultant reduction in protein expression was accompanied by attenuation of cAMP-repressive activity in the cells. We also demonstrate that human GPR176 is N-glycosylated. Importantly, missense variations in the conserved N-glycosylation sites of human GPR176 (rs1473415441; rs761894953) affected N-glycosylation and thereby attenuated protein expression and cAMP-repressive activity in the cells. We show that N-glycosylation is a prerequisite for the efficient protein expression of functional Gpr176/GPR176. |
著作権等: | © The Author(s) 2020. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
URI: | http://hdl.handle.net/2433/250077 |
DOI(出版社版): | 10.1038/s41598-020-61370-y |
PubMed ID: | 32157140 |
出現コレクション: | 学術雑誌掲載論文等 |
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