title: hCG-receptor binding and transmembrane signaling.
abstract: The technique of site-directed mutagenesis has proven to be quite powerful in elucidating contact sites involved in the interaction of the heterodimeric glycoprotein hormones and their respective seven transmembrane (TM) G protein-coupled receptors. Our laboratory has focused on identification of the minimum core sequences of the alpha and beta subunits required for bioactivity, the minimum length of a conjoined (yoked) single-chain hCG, the amino acid residues on hCG and the LH/CG-receptor (LH/CG-R) responsible for high-affinity binding, and the regions of the receptor that are involved in TM signaling. A number of amino acid residues have been mapped on the alpha and beta subunits of hCG that appear important in receptor binding. When projected onto the crystal structure of HF-treated hCG, these residues, by and large, cluster on one side of the molecule and cover a sizeable surface area, indicating that the hormone-receptor binding interface is rather extensive. Based on mutagenesis studies of several conserved ionizable amino acid residues in the extracellular domain (ECD) of LH/CG-R and a model that we, in collaboration with Drs Lapthorn and Isaacs, have developed for this region based on the crystal structure of porcine ribonuclease inhibitor, a charged region that appears to play an important role in hormone-receptor recognition has been identified. We have also delineated several regions of LH/CG-R that do not appear to participate in hCG binding but are involved in hCG-mediated signaling. These regions are located in the ECD and extracellular loop III just prior to entry into the membrane via TM helices I and VII, respectively, and in TM helices VI and VII. Similarly, a homologous region in the ECD of the FSH receptor, located with ten residues of TM helix I, is important in signaling but not hormone binding. These results suggest that ligand binding and ligand-mediated receptor activation are quasi-distinct, albeit sequential phenomena. Collectively, our mutagenesis and modeling studies, coupled with results from other laboratories, argue for a ligand-induced conformational change of the receptor that may involve a relative reorientation of the TM helices.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602, USA. puett@bchiris.biochem.uga.edu
authors.forename: D
authors.initials: D
authors.lastname: Puett
authors.forename: N
authors.initials: N
authors.lastname: Bhowmick
authors.forename: L M
authors.initials: LM
authors.lastname: Fernandez
authors.forename: J
authors.initials: J
authors.lastname: Huang
authors.forename: C
authors.initials: C
authors.lastname: Wu
authors.forename: P
authors.initials: P
authors.lastname: Narayan
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D006063:Chorionic Gonadotropin / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011974:Receptors, LH / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013329:Structure-Activity Relationship
chemical_list: D006063:Chorionic Gonadotropin; D011974:Receptors, LH