Cell-Surface Loss of Constitutive Activating and Inactivating Mutants of Eel Luteinizing Hormone Receptors
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6–6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.
Abstract
The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6–6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.
참고문헌 (32)
- Ascoli M, "The lutropin", choriogonadotropin receptor, a 2002 perspective, 2002, pp.23, 141~174
- Bhaskaran RS, "The post-endocytotic fate of the gonadotropin receptors is an important determinant of the desensitization of gonadotropin responses", J Mol Endocrinol, 34, pp.447~457 (2005)
- Byambaragchaa M, "Site specificity of eel luteinizing hormone N-linked oligosaccharides in signal transduction", Gen Comp Endocrinol, 268, pp.50~56 (2018)
- Munkhzaya Byambaragchaa, "Signal Transduction of Eel Luteinizing Hormone Receptor (eelLHR) and Follicle Stimulating Hormone Receptor (eelFSHR) by Recombinant Equine Chorionic Gonadotropin (rec-eCG) and Native eCG", 발생과 생식, 22(1), pp.55~64 (2018), 한국발생생물학회
- Byambaragchaa M, "Constitutive activation and inactivation of mutations inducing cell surface loss of receptor and impairing of signal transduction of agonist-stimulated eel follicle-stimulating hormone receptor", Int J Mol Sci, 21, pp.7075~ (2020)
- BYAMBARAGCHAA MUNKHZAYA, "Luteinizing hormone-like and follicle-stimulating hormone-like activities of equine chorionic gonadotropin β-subunit mutants in cells expressing rat luteinizing hormone", chorionic gonadotropin receptor and rat follicle-stimulating hormone receptor, 2021, pp.25(3), 171~181[DOI]
- Byambaragchaa M, "Constitutively activating mutants of equine LH", CGR constitutively induce signal transduction and inactivating mutations impair biological activity and cell-surface receptor loss in vitro, 2021, pp.22, 10723~
- Munkhzaya Byambaragchaa, "Constitutive Activating Eel Luteinizing Hormone Receptors Induce Constitutively Signal Transduction and Inactivating Mutants Impair Biological Activity", 발생과 생식, 25(3), pp.133~143 (2021), 한국발생생물학회
- BYAMBARAGCHAA MUNKHZAYA, "Functional characterization of naturally-occurring constitutively activating", inactivating mutations in equine follicle-stimulating hormone receptor, 2022, pp.35(3), 399~409[DOI]
- Dhanwada KR, "Two mutations of the lutropin", choriogonadotropin receptor that impair signal transduction also interfere with receptor-mediated endocytosis, 1996, pp.10, 544~554
- Foster SR, "Targeting trafficking in drug development. Handbook of Experimental Pharmacology" (2018), Springer
- Galet C, "A constitutively active mutant of the human lutropin receptor(hLHRL457R)escapes lysosomal targeting and degradation", Mol Endocrinol, 20, pp.2931~2945 (2006)
- Jacobsen SE, "The GPRC6A receptor displays constitutive internalization and sorting to the slow recycling pathway", J Biol Chem, 292, pp.6910~6926 (2017)
- Kim DJ, "Production and characterization of monoclonal antibodies against recombinant tethered follicle-stimulating hormone from Japanese eel Anguilla japonica", Gen Comp Endocrinol, 233, pp.8~15 (2016)
- Jeong-Min Kim, "The C-terminal Phosphorylation Sites of eel Follicle-Stimulating Hormone Receptor are Important Role in the Signal Transduction", 발생과 생식, 22(2), pp.143~153 (2018), 한국발생생물학회
- Kim JM, "Sitespecific roles of N-linked oligosaccharides in recombinant eel follicle-stimulating hormone for secretion and signal transduction", Gen Comp Endocrinol, 276, pp.37~44 (2019)
- Kudo M, "Transmembrane regions V and VI of the human luteinizing hormone receptor are required for constitutive activation by a mutation in the third intracellular loop", J Biol Chem, 271, pp.22470~22478 (1996)
- Latronico AC, "A homozygous microdeletion in helix 7 of the luteinizing hormone receptor associated with familial testicular and ovarian resistance is due to both decreased cell surface expression and impaired effector activation by the cell surface receptor", Mol Endocrinol, 12, pp.442~450 (1998)
- Latronico AC, "Insights learned from L457(3.43)R, an activating mutant of the human lutropin receptor", Mol Cell Endocrinol, 260-262, pp.287~293 (2007)
- Lee SY, "Roles of N-linked and O-linked glycosylation sites in the activity of equine chorionic gonadotropin in cells expressing rat luteinizing hormone", chorionic gonadotropin receptor and follicle-stimulating hormone receptor, 2021, pp.21, 52~
- McGee SR, "Precocious puberty and Leydig cell hyperplasia in male mice with a gain of function mutation in the LH receptor gene", Endocrinology, 154, pp.3900~3913 (2013)
- Meehan TP, "Constitutively active luteinizing hormone receptors: Consequences of in vivo expression", Mol Cell Endocrinol, 260-262, pp.294~300 (2007)
- Min KS, "Mutations that induce constitutive activation and mutations that impair signal transduction modulate the basal and", or agoniststimulated internalization of the lutropin, J Biol Chem, pp.1998, 273[DOI]
- Mundell SJ, "Deletion of the distal COOH-terminus of the A2B adenosine receptor switches internalization to an arrestin-and clathrin-independent pathway and inhibits recycling", Br J Pharmacol, 159, pp.518~533 (2010)
- Shenker A, "A constitutively activating mutation of the luteinizing hormone receptor in familial male precocious puberty", Nature, 365, pp.652~654 (1993)
- Simoni M, "The follicle-stimulating hormone receptor : Biochemistry, molecular biology, physiology, and pathophysiology", Endocr Rev, 18, pp.739~773 (1997)
- Tao YX, "Inactivating mutations of G protein-coupled receptors and diseases : Structurefunction insights and therapeutic implications", Pharmacol Ther, 111, pp.949~973 (2006)
- Tao YX, "Constitutive activation of G protein-coupled receptors as a result of selective substitution of a conserved leucine residue in transmembrane helix III", Mol Endocrinol, 14, pp.1272~1282 (2000)
- Themmen APN, "Mutations of gonadotropins and gonadotropin receptors : Elucidating the physiology and pathophysiology of pituitary-gonadal function", Endocr Rev, 21, pp.551~583 (2000)
- Yano K, "A case of male-limited precocious puberty caused by a point mutation in the second transmembrane domain of the luteinizing hormone choriogonadotropin receptor gene", Biochem Biophys Res Commun, 220, pp.1036~1042 (1996)
- Zhang M, "The formation of a salt bridge between helices 3 and 6 is responsible for the constitutive activity and lack of hormone responsiveness of the naturally occurring L457R mutation of the human lutropin receptor", J Biol Chem, 280, pp.26169~26176 (2005)
- Zhang M, "Intrinsic differences in the response of the human lutropin receptor versus the human follitropin receptor to activating mutations", J Biol Chem, 282, pp.25527~25539 (2007)
