The rate of internalization of the gonadotropin receptors is greatly affected by the origin of the extracellular domain.
Previous results from this laboratory have shown that human kidney (293) cells transfected with the rat follitropin receptor (rFSHR) internalize agonist (i.e. human follitropin, hFSH) at a rate similar to that of other agonist-G protein-coupled receptor complexes while 293 cells transfected with the rat lutropin/choriogonadotropin receptor (rLHR) internalize agonist (human choriogonadotropin, hCG) at a rate that is about 1 order of magnitude slower. Taking advantage of this difference and the high degree of homology between the rLHR and rFSHR, we have now used chimeras of these two receptors to begin to delineate structural features that influence their internalization. Analysis of six chimeras that exchanged only the transmembrane domains (designated FLF and LFL), only the COOH-terminal domains (FFL or LLF) or both domains (FLL or LFF) show that the origin of the extracellular domain is at least as important, if not more, than the origin of the transmembrane and COOH-terminal domains in determining the rate of internalization of the gonadotropin receptors. Thus, the rates of internalization of agonist internalization mediated by FFL, FLF, and FLL more closely resemble rFSHR than rLHR, while the rates of agonist internalization mediated by LLF, LFL, and LFF more closely resemble rLHR than rFSHR. The importance of the extracellular domain was also evident even upon overexpression of arrestin-3, a protein that enhances the rate of internalization of the wild-type receptors and chimeras by binding to their intracellular regions.
Nakamura
K
K
Department of Pharmacology, The University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
Liu
X
X
Ascoli
M
M
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D051381:Rats; D011967:Receptors, Gonadotropin / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D017421:Sequence Analysis; D015398:Signal Transduction
D006063:Chorionic Gonadotropin; D011967:Receptors, Gonadotropin; D011993:Recombinant Fusion Proteins; D019204:GTP-Binding Proteins