Reconstitution of a high-affinity functional lutropin receptor by coexpression of its extracellular and membrane domains.
The glycoprotein hormone receptors differ from other G protein-coupled receptors by their large extracellular domain which mediates ligand binding. Cooperation between the G-protein coupled membrane domain, the extracellular domain and the hormone in establishing high-affinity binding and efficient transduction is likely to exist. Expression plasmids encoding the full-length porcine LH-hCG receptor (1-696), its extracellular (1-297) and membrane domain (298-696), as well as the alpha and beta subunits of hCG were constructed. We report that coexpression in COS cells of the two LH-hCG receptor domains restores cell surface high-affinity hormone binding and hormone dependent adenylyl cyclase activation, suggesting sufficient interactions between the two receptor domains to reconstitute a complete functional molecule. Moreover, the two hormone subunits and the two receptor domains are able to associate within coexpressing COS cells into an active complex.
Remy
J J
JJ
Unité d'Ingénierie des Protéines, INRA-Biotechnologies, Jouy-en-Josas, France.
Bozon
V
V
Couture
L
L
Goxe
B
B
Salesse
R
R
Garnier
J
J
D000262:Adenylyl Cyclases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D006063:Chorionic Gonadotropin / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism; D007668:Kidney; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D011974:Receptors, LH / Q000096:biosynthesis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013552:Swine; D014162:Transfection
D006063:Chorionic Gonadotropin; D046911:Macromolecular Substances; D011974:Receptors, LH; D011994:Recombinant Proteins; D004247:DNA; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases