Receptor activation of and signal generation by the lutropin/choriogonadotropin receptor. Cooperation of Asp397 of the receptor and alpha Lys91 of the hormone.
We have developed a novel method of reciprocal substitution mutation to identify pairing of amino acids within a receptor and its ligand. Using this method, we demonstrate for the first time that a pair of counterionic amino acids, one from the lutropin/choriogonadotropin receptor and the other from the ligand (human choriogonadotropin), cooperate and perhaps interact with each other to activate the receptor and to generate hormonal signal. In this study, Asp397 of the receptor was converted to Lys while Lys91 of the alpha subunit of human choriogonadotropin was substituted with Asp, thus maintaining the counterionic nature of this pair of amino acids. Mutation at each of these positions does not affect the hormone-receptor interaction but results in the significant or complete loss of the bioactivity of both the receptor and the hormone. However, when the impotent mutant receptor and mutant hormone were paired to interact together, they induced cAMP synthesis, resulting in a potent receptor-hormone couple. Substitutions with other amino acids that eliminated the counterionic nature failed to induce cAMP synthesis. Our results shed light on the molecular mechanisms of receptor activation and will serve as a model for other G-protein-coupled peptide receptors, particularly glycoprotein hormone receptors.
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Department of Molecular Biology, University of Wyoming, Laramie 82071-3944.
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D001224:Aspartic Acid / Q000737:chemistry; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000737:chemistry / Q000378:metabolism*; D000242:Cyclic AMP / Q000096:biosynthesis; D019204:GTP-Binding Proteins / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D007986:Luteinizing Hormone / Q000737:chemistry / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D011974:Receptors, LH / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D014162:Transfection
D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D011994:Recombinant Proteins; D001224:Aspartic Acid; D007986:Luteinizing Hormone; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D008239:Lysine