Mutations that induce constitutive activation and mutations that impair signal transduction modulate the basal and/or agonist-stimulated internalization of the Lutropin/Choriogonadotropin receptor.
Previous results from this laboratory suggested that the same active conformation of the lutropin/choriogonadotropin receptor (LHR) is involved in the stimulation of G proteins and in triggering the internalization of the bound agonist. We have now analyzed two naturally occurring, constitutively active mutants of the human LHR. These mutations were introduced into the rat LHR (rLHR) and are designated L435R and D556Y. Cells expressing rLHR-D556Y bind human choriogonadotropin (hCG) with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation does not affect the internalization of the free receptor, but it enhances the internalization of the agonist-occupied receptors approximately 3-fold. Cells expressing rLHR-L435R also bind hCG with normal affinity, exhibit a 47-fold increase in basal cAMP, and do not respond to hCG with a further increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors approximately 2- and approximately 17-fold, respectively. We conclude that the state of activation of the rLHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing rLHR-L435R is due to the fast rate of internalization of the bound hCG. The finding that membranes expressing rLHR-L435R (a system where internalization does not occur) respond to hCG with an increase in adenylyl cyclase activity supports this suggestion.
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Department of Pharmacology, University of Iowa College of Medicine, Iowa City, Iowa 52242-1109, USA. mario-ascoli@uiowa.edu
Liu
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Fabritz
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Jaquette
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Abell
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Ascoli
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D000262:Adenylyl Cyclases / Q000032:analysis; D000818:Animals; D001692:Biological Transport; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D002999:Clone Cells; D004705:Endocytosis; D006801:Humans; D007700:Kinetics; D007986:Luteinizing Hormone / Q000378:metabolism*; D009154:Mutation; D051381:Rats; D011974:Receptors, LH / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction
D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D011994:Recombinant Proteins; D007986:Luteinizing Hormone; D000262:Adenylyl Cyclases