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Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor

Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits on...

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Autores principales: Byambaragchaa, Munkhzaya, Choi, Seung-Hee, Joo, Hyo-Eun, Kim, Sang-Gwon, Kim, Yean-Ji, Park, Gyeong-Eun, Kang, Myung-Hwa, Min, Kwan-Sik
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society of Developmental Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8807129/
https://www.ncbi.nlm.nih.gov/pubmed/35141446
http://dx.doi.org/10.12717/DR.2021.25.4.199
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author Byambaragchaa, Munkhzaya
Choi, Seung-Hee
Joo, Hyo-Eun
Kim, Sang-Gwon
Kim, Yean-Ji
Park, Gyeong-Eun
Kang, Myung-Hwa
Min, Kwan-Sik
author_facet Byambaragchaa, Munkhzaya
Choi, Seung-Hee
Joo, Hyo-Eun
Kim, Sang-Gwon
Kim, Yean-Ji
Park, Gyeong-Eun
Kang, Myung-Hwa
Min, Kwan-Sik
author_sort Byambaragchaa, Munkhzaya
collection PubMed
description Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits only LH-like activity. To identify the specific biological functions of glycosylated sites in eCG, we constructed the following site mutants of N- and O-linked glycosylation: eCGβ/αΔ56, substitution of α-subunit(56) N-linked glycosylation site; eCGβ-D/α, deletion of the O-linked glycosylation sites at the β-subunit, and eCGβ-D/αΔ56, double mutant. We produced recombinant eCG (rec-eCG) proteins in Chinese hamster ovary suspension (CHO-S) cells. We examined the biological activity of rec-eCG proteins in CHO-K1 cells expressing the eLH/CG receptor and found that signal transduction activities of deglycosylated mutants remarkably decreased. The EC(50) levels of eCGβ/αΔ56, eCGβ-D/α, and eCGβ-D/αΔ56 mutants decreased by 2.1-, 5.6-, and 3.4-fold, respectively, compared to that of wild-type eCG. The Rmax values of the mutants were 56%-80% those of wild-type eCG (141.9 nmol/10(4) cells). Our results indicate that the biological activity of eCG is greatly affected by the removal of N- and O-linked glycosylation sites in cells expressing eLH/CGR. These results provide important information on rec-eCG in the regulation of specific glycosylation sites and improve our understanding of the specific biological activity of rec-eCG glycosylation sites in equidaes.
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spelling pubmed-88071292022-02-08 Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor Byambaragchaa, Munkhzaya Choi, Seung-Hee Joo, Hyo-Eun Kim, Sang-Gwon Kim, Yean-Ji Park, Gyeong-Eun Kang, Myung-Hwa Min, Kwan-Sik Dev Reprod Research Article Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits only LH-like activity. To identify the specific biological functions of glycosylated sites in eCG, we constructed the following site mutants of N- and O-linked glycosylation: eCGβ/αΔ56, substitution of α-subunit(56) N-linked glycosylation site; eCGβ-D/α, deletion of the O-linked glycosylation sites at the β-subunit, and eCGβ-D/αΔ56, double mutant. We produced recombinant eCG (rec-eCG) proteins in Chinese hamster ovary suspension (CHO-S) cells. We examined the biological activity of rec-eCG proteins in CHO-K1 cells expressing the eLH/CG receptor and found that signal transduction activities of deglycosylated mutants remarkably decreased. The EC(50) levels of eCGβ/αΔ56, eCGβ-D/α, and eCGβ-D/αΔ56 mutants decreased by 2.1-, 5.6-, and 3.4-fold, respectively, compared to that of wild-type eCG. The Rmax values of the mutants were 56%-80% those of wild-type eCG (141.9 nmol/10(4) cells). Our results indicate that the biological activity of eCG is greatly affected by the removal of N- and O-linked glycosylation sites in cells expressing eLH/CGR. These results provide important information on rec-eCG in the regulation of specific glycosylation sites and improve our understanding of the specific biological activity of rec-eCG glycosylation sites in equidaes. Korean Society of Developmental Biology 2021-12 2021-12-31 /pmc/articles/PMC8807129/ /pubmed/35141446 http://dx.doi.org/10.12717/DR.2021.25.4.199 Text en © Copyright 2021 The Korean Society of Developmental Biology https://creativecommons.org/licenses/by-nc/3.0/This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creative-commons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Byambaragchaa, Munkhzaya
Choi, Seung-Hee
Joo, Hyo-Eun
Kim, Sang-Gwon
Kim, Yean-Ji
Park, Gyeong-Eun
Kang, Myung-Hwa
Min, Kwan-Sik
Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title_full Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title_fullStr Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title_full_unstemmed Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title_short Specific Biological Activity of Equine Chorionic Gonadotropin (eCG) Glycosylation Sites in Cells Expressing Equine Luteinizing Hormone/CG (eLH/CG) Receptor
title_sort specific biological activity of equine chorionic gonadotropin (ecg) glycosylation sites in cells expressing equine luteinizing hormone/cg (elh/cg) receptor
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8807129/
https://www.ncbi.nlm.nih.gov/pubmed/35141446
http://dx.doi.org/10.12717/DR.2021.25.4.199
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