Constitutive Activating Eel Luteinizing Hormone Receptors Induce Constitutively Signal Transduction and Inactivating Mutants Impair Biological Activity
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In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.
Abstract
In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.
참고문헌 (28)
- Althumairy D, "Glycoprotein G-protein coupled receptors in disease: Luteinizing hormone receptors and follicle stimulating hormone receptors", Diseases, 8, pp.35~ (2020)
- Ascoli M, "The lutropin", choriogonadotropin gonadotropin receptor, a 2002 perspective, 2002, pp.23, 141~174
- Byambaragchaa M, "Site specificity of eel luteinizing hormone N-linked oligosaccharides in signal transduction", Gen Comp Endocrinol, 268, pp.50~56 (2018)
- 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)
- Dhanwada KR, "Two mutations of the lutropin", choriogonadotropin receptor that impair signal transduction also interfere with receptor-mediated endocytosis, 1996, pp.10, 544~554
- Galet C, "A constitutively active mutant of the human lutropin receptor(hLHRL457R)escapes lysosomal targeting and degradation", Mol Endocrinol, 20, pp.2931~2945 (2006)
- Huhtaniemi I, "Activating and inactivating hormone receptor mutations", Horm Res, 53, pp.S9~S16 (2000)
- Huhtaniemi IT, "Mutations in human gonadotropin and gonadotropinreceptor genes", Endocrine, 26, pp.207~217 (2005)
- Ignacak M, "Substitution of M398T in the second transmembrane helix of the LH receptor in a patient with familial male-limited precocious puberty", Endocr J, 47, pp.595~599 (2000)
- 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)
- Kim JM, "Site-specific roles of N-linked oligosaccharides in recombinant eel follicle-stimulating hormone for secretion and signal transduction", Gen Comp Endocrinol, 276, pp.37~44 (2019)
- Kraaij R, "A missense mutation in the second transmembrane segment of the luteinizing hormone receptor causes familial male-limited precocious puberty", J Clin Endocrinol Metab, 80, pp.3168~3172 (1995)
- 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, pp.260~262 (2007)
- 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]
- Quintana J, "The regulation of the binding affinity of the luteinizing hormone", choriogonadotropin receptor by sodium ions is mediated by a highly conserved aspartate located in the second transmembrane domain of G protein-coupled receptors, 1993, pp.7, 767~775
- Segaloff DL, "The lutropin", choriogonadotropin receptor… 4 years later, 1993, pp.14, 324~347
- Shenker A, "Activating mutations of the lutropin choriogonadotropin receptor in precocious puberty", Recept Channels, 8, pp.3~18 (2002)
- Shenker A, "A constitutively activating mutation of the luteinizing hormone receptor in familial male precocious puberty", Nature, 365, pp.652~654 (1993)
- 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)
- Wu SM, "Luteinizing hormone receptor mutations in disorders of sexual development and cancer", Front Biosci, 5, pp.D343~D352 (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)
