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Signal Transduction of Constitutive Activating and Inactivating Eel Lutropin/ Choriogonadotropin Receptor (Eel LH/ CGR) Mutants by Recombinant Equine Chorionic Gonadotropin (Rec-eCG)

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Lutropin/choriogonadotropin receptor (LH/CGR) is a member of the G protein-coupled receptor superfamily. LH/CGRs in fish and mammalian species have been reported to contain naturally occurring, constitutively activating, and inactivating mutations in highly conserved regions. The present study was designed to determine the functional aspect of eel LH/CGR signal transduction. Biochemical analysis was performed using cells transfected with wild-type eel LH/CG (eel LH/CGR-wt) or with activating (designated eel LH/CGR-M410T, L469R, and D590Y) and inactivating (eel LH/CGR-D 417N and Y558F) mutants. We also generated a mutant (eel LH/CGR-t651) in which the C-terminal cytoplasmic tail was truncated at residue 651. Activating mutant cells expressing eel LH/CGR-M410T, L469R, and D590Y exhibited 1.4-, 8.7-, and 4.0-fold increases in the basal cAMP response, respectively, without recombinant equine chorionic gonadotropin (rec-eCG) agonist treatment. In inactivating mutants (eel LH/CGR-D417N and Y558F), the cyclic adenosine monophosphate (cAMP) response did not result in completely impaired signal transduction. However, the eel LH/ CGR-t651 mutant did not exhibit any cAMP signaling following high-agonist treatment. Rmax values did not increase with further rec-eCG agonist stimulation. Our results suggest that constitutively activating and inactivating eel LH/CGR mutants with highly conserved amino acids exhibit a significant signal transduction pathway for glycoprotein hormone receptors. Eel LH/CGRs in activating and inactivating mutants are usually processed by receptor-mediated signaling following rec-eCG agonist stimulation.

Abstract

Lutropin/choriogonadotropin receptor (LH/CGR) is a member of the G protein-coupled receptor superfamily. LH/CGRs in fish and mammalian species have been reported to contain naturally occurring, constitutively activating, and inactivating mutations in highly conserved regions. The present study was designed to determine the functional aspect of eel LH/CGR signal transduction. Biochemical analysis was performed using cells transfected with wild-type eel LH/CG (eel LH/CGR-wt) or with activating (designated eel LH/CGR-M410T, L469R, and D590Y) and inactivating (eel LH/CGR-D 417N and Y558F) mutants. We also generated a mutant (eel LH/CGR-t651) in which the C-terminal cytoplasmic tail was truncated at residue 651. Activating mutant cells expressing eel LH/CGR-M410T, L469R, and D590Y exhibited 1.4-, 8.7-, and 4.0-fold increases in the basal cAMP response, respectively, without recombinant equine chorionic gonadotropin (rec-eCG) agonist treatment. In inactivating mutants (eel LH/CGR-D417N and Y558F), the cyclic adenosine monophosphate (cAMP) response did not result in completely impaired signal transduction. However, the eel LH/ CGR-t651 mutant did not exhibit any cAMP signaling following high-agonist treatment. Rmax values did not increase with further rec-eCG agonist stimulation. Our results suggest that constitutively activating and inactivating eel LH/CGR mutants with highly conserved amino acids exhibit a significant signal transduction pathway for glycoprotein hormone receptors. Eel LH/CGRs in activating and inactivating mutants are usually processed by receptor-mediated signaling following rec-eCG agonist stimulation.

참고문헌 (34)

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관련 논문AI추천

Constitutive Activating Eel Luteinizing Hormone Receptors Induce Constitutively Signal Transduction and Inactivating Mutants Impair Biological Activity민관식, Dong-Wan Kim, Munkhzaya Byambaragchaa, Seung-Hee Choi한국발생생물학회 · 발생과 생식2021 · 제25권 제3호Cell-Surface Loss of Constitutive Activating and Inactivating Mutants of Eel Luteinizing Hormone Receptors민관식, Dong-Wan Kim1, Munkhzaya Byambaragchaa, Seung-Hee Choi한국발생생물학회 · 발생과 생식2021 · 제25권 제4호The C-terminal Phosphorylation Sites of eel Follicle-Stimulating Hormone Receptor are Important Role in the Signal Transduction강명화, 민관식, Jeong-Min Kim, Munkhzaya Byambaragchaa한국발생생물학회 · 발생과 생식2018 · 제22권 제2호Signal Transduction of Eel Luteinizing Hormone Receptor (eelLHR) and Follicle Stimulating Hormone Receptor (eelFSHR) by Recombinant Equine Chorionic Gonadotropin (rec-eCG) and Native eCG강명화, 민관식, Dae-Jung Kim, Munkhzaya Byambaragchaa, So-Yun Lee한국발생생물학회 · 발생과 생식2018 · 제22권 제1호Signal Transduction of C-Terminal Phosphorylation Regions for Equine Luteinizing Hormone/Chorionic Gonadotropin Receptor (eLH/CGR)민관식한국발생생물학회 · 발생과 생식2022 · 제26권 제1호Functional characterization of naturally-occurring constitutively activating/inactivating mutations in equine follicle-stimulating hormone receptor민관식, Ahn Tae-Young, BYAMBARAGCHAA MUNKHZAYA, Choi Seung-Hee, Kang Myung-Hwa아세아·태평양축산학회 · Animal Bioscience2022 · 제35권 제3호

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