title: Asp330 and Tyr331 in the C-terminal cysteine-rich region of the luteinizing hormone receptor are key residues in hormone-induced receptor activation.
abstract: The luteinizing hormone (LH) receptor plays an essential role in male and female gonadal function. Together with the follicle-stimulating hormone (FSH) and thyroid stimulating hormone (TSH) receptors, the LH receptor forms the family of glycoprotein hormone receptors. All glycoprotein hormone receptors share a common modular topography, with an N-terminal extracellular ligand binding domain and a C-terminal seven-transmembrane transduction domain. The ligand binding domain consists of 9 leucine-rich repeats, flanked by N- and C-terminal cysteine-rich regions. Recently, crystal structures have been published of the extracellular domains of the FSH and TSH receptors. However, the C-terminal cysteine-rich region (CCR), also referred to as the "hinge region," was not included in these structures. Both structure and function of the CCR therefore remain unknown. In this study we set out to characterize important domains within the CCR of the LH receptor. First, we mutated all cysteines and combinations of cysteines in the CCR to identify the most probable disulfide bridges. Second, we exchanged large parts of the LH receptor CCR by its FSH receptor counterparts, and characterized the mutant receptors in transiently transfected HEK 293 cells. We zoomed in on important regions by focused exchange and deletion mutagenesis followed by alanine scanning. Mutations in the CCR specifically decreased the potencies of LH and hCG, because the potency of the low molecular weight agonist Org 41841 was unaffected. Using this unbiased approach, we identified Asp(330) and Tyr(331) as key amino acids in LH/hCG mediated signaling.
authors.affiliation: Department of Internal Medicine, Erasmus MC, P. O. Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Bruysters
authors.forename: Miriam
authors.initials: M
authors.lastname: Verhoef-Post
authors.forename: Axel P N
authors.initials: AP
authors.lastname: Themmen
mesh_terms: D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000737:chemistry*; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides; D017930:Genes, Reporter; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011962:Receptors, FSH / Q000378:metabolism; D011974:Receptors, LH / Q000737:chemistry* / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D004220:Disulfides; D011962:Receptors, FSH; D011974:Receptors, LH; D001224:Aspartic Acid; D014443:Tyrosine; D003545:Cysteine