{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T20:20:25Z","timestamp":1761942025835,"version":"build-2065373602"},"reference-count":52,"publisher":"The Endocrine Society","issue":"11","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,11,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Using biochemical and imaging approaches, we examined the postendocytotic fate of the complex formed by human choriogonadotropin (hCG) and a constitutively active mutant of the human lutropin receptor (hLHR-L457R) found in a boy with precocious puberty and Leydig cell hyperplasia. After internalization, some of the complex formed by the hLHR-wild type (hLHR-wt) and hCG recycles to the cell surface, and some is found in lysosomes where the hormone is degraded. In contrast, the complex formed by the hLHR-L457R and hCG is not routed to the lysosomes, most of it is recycled to the cell surface and hormone degradation is barely detectable. For both, hLHR-wt and -L457R, there is an hCG-induced loss of cell surface receptors that accompanies internalization but this loss cannot be prevented by leupeptin. The removal of recycling motifs of the hLHR by truncation of the C-terminal tail at residue 682 greatly enhances the lysosomal accumulation of the hormone-receptor complexes formed by the hLHR-wt or the L457R mutant, the degradation of the internalized hormone, and the loss of cell surface receptors. The degradation of the hormone internalized by these mutants as well as the loss of cell surface receptors is largely prevented by leupeptin. These results highlight a previously unrecognized complexity in the postendocytotic trafficking of the hLHR and document a clear difference between the properties of the constitutively active mutant and the agonist-activated hLHR-wt. This lack of lysosomal degradation of the L457R mutant could contribute to its constitutive activity by prolonging the duration of signaling.<\/jats:p>","DOI":"10.1210\/me.2006-0138","type":"journal-article","created":{"date-parts":[[2006,6,28]],"date-time":"2006-06-28T01:27:31Z","timestamp":1151458051000},"page":"2931-2945","source":"Crossref","is-referenced-by-count":13,"title":["A Constitutively Active Mutant of the Human Lutropin Receptor (hLHR-L457R) Escapes Lysosomal Targeting and Degradation"],"prefix":"10.1210","volume":"20","author":[{"given":"Colette","family":"Galet","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Roy J. and Lucille A. Carver College of Medicine, The University of Iowa, Iowa City, Iowa 52242-1109"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mario","family":"Ascoli","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Roy J. and Lucille A. Carver College of Medicine, The University of Iowa, Iowa City, Iowa 52242-1109"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"80","reference":[{"key":"2020071613564708800_R1","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1038\/35094577","article-title":"Classical and new roles of \u03b2-arrestins in the regulation of G-protein-coupled receptors.","volume":"2","author":"Pierce","year":"2001","journal-title":"Nat Rev Neurosci"},{"key":"2020071613564708800_R2","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/S0955-0674(00)00190-3","article-title":"Expanding roles for \u03b2-arrestins as scaffolds and adapters in GPCR signaling and trafficking.","volume":"13","author":"Miller","year":"2001","journal-title":"Curr Opin Cell Biol"},{"key":"2020071613564708800_R3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0031-6997(24)01478-9","article-title":"Evolving concepts in G protein-coupled receptor endocytosis: the role in receptor desensitization and signaling.","volume":"53","author":"Ferguson","year":"2001","journal-title":"Pharmacol Rev"},{"key":"2020071613564708800_R4","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/S0962-8924(01)02239-5","article-title":"Arresting developments in heptahelical receptor signaling and regulation.","volume":"12","author":"Perry","year":"2002","journal-title":"Trends Cell Biol"},{"key":"2020071613564708800_R5","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1038\/nrm883","article-title":"Signal transduction and endocytosis: close encounters of many kinds.","volume":"3","author":"Sorkin","year":"2002","journal-title":"Nat Rev Mol Cell Biol"},{"key":"2020071613564708800_R6","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/S0968-0004(03)00134-8","article-title":"The ins and outs of G protein-coupled receptor trafficking.","volume":"28","author":"Marchese","year":"2003","journal-title":"Trends Biochem Sci"},{"key":"2020071613564708800_R7","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/S0165-6147(00)01620-5","article-title":"Role of endocytosis in mediating downregulation of G-protein-coupled receptors.","volume":"22","author":"Tsao","year":"2001","journal-title":"Trends Pharmacol Sci"},{"key":"2020071613564708800_R8","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1038\/35056583","article-title":"Protein regulation by monoubiquitin.","volume":"2","author":"Hicke","year":"2001","journal-title":"Nat Rev Mol Cell Biol"},{"key":"2020071613564708800_R9","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1016\/S0092-8674(01)00485-8","article-title":"A new ticket for entry into budding vesicles-ubiquitin.","volume":"106","author":"Hicke","year":"2001","journal-title":"Cell"},{"key":"2020071613564708800_R10","doi-asserted-by":"crossref","first-page":"1242","DOI":"10.1083\/jcb.99.4.1242","article-title":"Lysosomal accumulation of the hormone-receptor complex during receptor-mediated endocytosis of human choriogonadotropin.","volume":"99","author":"Ascoli","year":"1984","journal-title":"J Cell Biol"},{"key":"2020071613564708800_R11","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1016\/S0021-9258(17)35240-7","article-title":"Lutropin\/choriogonadotropin down-regulates its receptor by both receptor mediated endocytosis and a cAMP-dependent reduction in receptor mRNA.","volume":"266","author":"Wang","year":"1991","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R12","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1677\/jme.1.01745","article-title":"The post-endocytotic fate of the gonadotropin receptors is an important determinant of the desensitization of gonadotropin responses.","volume":"34","author":"Bhaskaran","year":"2005","journal-title":"J Mol Endocrinol"},{"key":"2020071613564708800_R13","doi-asserted-by":"crossref","first-page":"13698","DOI":"10.1073\/pnas.95.23.13698","article-title":"Termination of signaling by protease-activated receptor-1 is linked to lysosomal sorting.","volume":"95","author":"Trejo","year":"1998","journal-title":"Proc Natl Acad Sci USA"},{"key":"2020071613564708800_R14","doi-asserted-by":"crossref","first-page":"2216","DOI":"10.1074\/jbc.274.4.2216","article-title":"The cytoplasmic tails of protease-activated receptor-1 and substance P receptor specify sorting to lysosomes versus recycling.","volume":"274","author":"Trejo","year":"1999","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R15","doi-asserted-by":"crossref","first-page":"17596","DOI":"10.1074\/jbc.M000142200","article-title":"Regulation and intracellular trafficking pathways of the endothelin receptors.","volume":"275","author":"Bremnes","year":"2000","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R16","doi-asserted-by":"crossref","first-page":"1624","DOI":"10.1210\/mend.15.9.0698","article-title":"Identification of two distinct structural motifs that, when added to the C-terminal tail of the rat lutropin receptor, redirect the internalized hormone-receptor complex from a degradation to a recycling pathway.","volume":"15","author":"Kishi","year":"2001","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R17","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1210\/me.2002-0161","article-title":"Identification of a transferable two amino acid motif (GT) present in the C-terminal tail of the human lutropin receptor that redirects internalized G protein-coupled receptors from a degradation to a recycling pathway.","volume":"17","author":"Galet","year":"2003","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R18","doi-asserted-by":"crossref","first-page":"49348","DOI":"10.1074\/jbc.M306557200","article-title":"GIPC binds to the human lutropin receptor (LHR) through an unusual PDZ domain binding motif and it regulates the sorting of the internalized human choriogonadotropin (hCG) and the density of cell surface LHR.","volume":"278","author":"Hirakawa","year":"2003","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R19","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1210\/me.2003-0293","article-title":"The postendocytotic trafficking of the hLHR is mediated by a transferable motif consisting of the C-terminal cysteine and an upstream leucine.","volume":"18","author":"Galet","year":"2004","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R20","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1210\/edrv.23.2.0462","article-title":"The lutropin\/choriogonadotropin receptor. A 2002 perspective.","volume":"23","author":"Ascoli","year":"2002","journal-title":"Endocr Rev"},{"key":"2020071613564708800_R21","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1530\/rep.1.00663","article-title":"An update of the pathophysiology of human gonadotrophin subunit and receptor gene mutations and polymorphisms.","volume":"130","author":"Themmen","year":"2005","journal-title":"Reproduction"},{"key":"2020071613564708800_R22","doi-asserted-by":"crossref","first-page":"34911","DOI":"10.1074\/jbc.273.52.34911","article-title":"Mutations that induce constitutive activation and mutations that impair signal transduction modulate the basal and\/or agonist-stimulated internalization of the lutropin\/choriogonadotropin receptor.","volume":"273","author":"Min","year":"1998","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R23","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1210\/mend.14.11.0555","article-title":"Effect of activating and inactivating mutations on the phosphorylation and trafficking of the human lutropin\/choriogonadotropin receptor.","volume":"14","author":"Min","year":"2000","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R24","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1074\/jbc.M106082200","article-title":"The association of arrestin-3 with the human lutropin\/choriogonadotropin receptor depends mostly on receptor activation rather than on receptor phosphorylation.","volume":"277","author":"Min","year":"2002","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R25","doi-asserted-by":"crossref","first-page":"8703","DOI":"10.1021\/bi990169t","article-title":"Evidence that constitutively active luteinizing hormone\/human chorionic gonadotropin receptors are rapidly internalized.","volume":"38","author":"Bradbury","year":"1999","journal-title":"Biochemistry"},{"key":"2020071613564708800_R26","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1210\/me.2004-0335","article-title":"Evidence that palmitoylation of carboxyl terminus cysteine residues of the human luteinizing hormone receptor regulates postendocytic processing.","volume":"19","author":"Munshi","year":"2005","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R27","first-page":"2435","article-title":"A unique constitutively activating mutation in third transmbembrane helix of luteinizing hormone receptor causes sporadic male gonadotropin-independent precocious puberty.","volume":"83","author":"Latronico","year":"1998","journal-title":"J Clin Endocrinol Metab"},{"key":"2020071613564708800_R28","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1210\/mend.15.6.0661","article-title":"Pleiotropic effects of substitutions of a highly conserved leucine in transmembrane helix III of the human lutropin\/choriogonadotropin receptor with respect to constitutive activation and hormone responsiveness.","volume":"15","author":"Shinozaki","year":"2001","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R29","doi-asserted-by":"crossref","first-page":"3872","DOI":"10.1210\/en.2003-0365","article-title":"A constitutively active somatic mutation of the human lutropin receptor found in Leydig cell tumors activates the same families of G proteins as germ line mutations associated with Leydig cell hyperplasia.","volume":"144","author":"Hirakawa","year":"2003","journal-title":"Endocrinology"},{"key":"2020071613564708800_R30","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1210\/endo.143.3.8702","article-title":"MA-10 cells transfected with the human lutropin\/choriogonadotropin receptor (hLHR). A novel experimental paradigm to study the functional properties of the hLHR.","volume":"143","author":"Hirakawa","year":"2002","journal-title":"Endocrinology"},{"key":"2020071613564708800_R31","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1210\/mend.13.8.0331","article-title":"Role of the rate of internalization of the agonist-receptor complex on the agonist-induced down-regulation of the lutropin\/choriogonadotropin receptor.","volume":"13","author":"Nakamura","year":"1999","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R32","doi-asserted-by":"crossref","first-page":"25426","DOI":"10.1074\/jbc.274.36.25426","article-title":"The rate of internalization of the gonadotropin receptors is greatly affected by the origin of the extracellular domain.","volume":"274","author":"Nakamura","year":"1999","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R33","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1016\/S0026-895X(24)12534-5","article-title":"A dileucine-based motif in the C-terminal tail of the lutropin\/choriogonadotropin receptor inhibits endocytosis of the agonist-receptor complex.","volume":"56","author":"Nakamura","year":"1999","journal-title":"Mol Pharmacol"},{"key":"2020071613564708800_R34","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1074\/jbc.275.1.241","article-title":"Seven non-contiguous intracellular residues of the lutropin\/choriogonadotropin receptor dictate the rate of agonist-induced internalization and its sensitivity to non-visual arrestins.","volume":"275","author":"Nakamura","year":"2000","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R35","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1210\/mend.14.6.0475","article-title":"The C-terminal tail of the rat lutropin\/choriogonadotropin receptor independently modulates hCG-induced internalization of the cell surface receptor and the lysosomal targeting of the internalized hCG-receptor complex.","volume":"14","author":"Kishi","year":"2000","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R36","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1083\/jcb.118.6.1347","article-title":"Pathways of internalization of the hCG\/LH receptor: immunoelectron microscopic studies in Leydig cells and transfected L cells.","volume":"118","author":"Ghinea","year":"1992","journal-title":"J Cell Biol"},{"key":"2020071613564708800_R37","doi-asserted-by":"crossref","first-page":"1751","DOI":"10.1210\/mend.13.10.0360","article-title":"Internalization and recycling pathways of the thyrotropin receptor.","volume":"13","author":"Baratti-Elbaz","year":"1999","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R38","doi-asserted-by":"crossref","first-page":"2809","DOI":"10.1083\/jcb.109.6.2809","article-title":"Polarized endocytosis by Madin-Darby canine kidney cells transfected with functional chicken liver glycoprotein receptor.","volume":"109","author":"Graeve","year":"1989","journal-title":"J Cell Biol"},{"key":"2020071613564708800_R39","doi-asserted-by":"crossref","first-page":"44712","DOI":"10.1074\/jbc.M107417200","article-title":"A transplantable sorting signal that is sufficient to mediate rapid recycling of G protein-coupled receptors.","volume":"276","author":"Gage","year":"2001","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R40","doi-asserted-by":"crossref","first-page":"45978","DOI":"10.1074\/jbc.M304504200","article-title":"A novel endocytic recycling signal that distinguishes the membrane trafficking of naturally occurring opioid receptors.","volume":"278","author":"Tanowitz","year":"2003","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R41","doi-asserted-by":"crossref","first-page":"7832","DOI":"10.1016\/S0021-9258(17)34446-0","article-title":"Inhibition of the degradation of receptor-bound human choriogonadotropin by lysosomotropic agents, protease inhibitors, and metabolic inhibitors.","volume":"253","author":"Ascoli","year":"1978","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R42","doi-asserted-by":"crossref","first-page":"1581","DOI":"10.1124\/mol.104.007013","article-title":"Subtype-specific sorting of the ETA endothelin receptor by a novel endocytic recycling signal for G protein-coupled receptors.","volume":"67","author":"Paasche","year":"2005","journal-title":"Mol Pharmacol"},{"key":"2020071613564708800_R43","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1016\/j.tem.2004.06.009","article-title":"Intracellular trafficking of hormone receptors.","volume":"15","author":"Gaborik","year":"2004","journal-title":"Trends Endocrinol Metab"},{"key":"2020071613564708800_R44","first-page":"767","article-title":"The regulation of the binding affinity of the luteinizing hormone\/choriogonadotropin receptor by sodium ions is mediated by a highly conserved aspartate located in the second transmembrane domain of G protein-coupled receptors.","volume":"7","author":"Quintana","year":"1993","journal-title":"Mol Endocrinol"},{"key":"2020071613564708800_R45","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1530\/rep.1.00662","article-title":"Specificity and promiscuity of gonadotropin receptors.","volume":"130","author":"Costagliola","year":"2005","journal-title":"Reproduction"},{"key":"2020071613564708800_R46","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1021\/bi972355+","article-title":"Transfected cells express mostly the intracellular precursor of the lutropin\/choriogonadotropin receptor but this precursor binds choriogonadotropin with high affinity.","volume":"37","author":"Fabritz","year":"1998","journal-title":"Biochemistry"},{"key":"2020071613564708800_R47","doi-asserted-by":"crossref","first-page":"2745","DOI":"10.1128\/MCB.7.8.2745","article-title":"High-efficiency transformation of mammalian cells by plasmid DNA.","volume":"7","author":"Chen","year":"1987","journal-title":"Mol Cell Biol"},{"key":"2020071613564708800_R48","doi-asserted-by":"crossref","first-page":"13306","DOI":"10.1016\/S0021-9258(18)33447-1","article-title":"Internalization and degradation of receptor-bound human choriogonadotropin in Leydig tumor cells. Fate of the hormone subunits.","volume":"257","author":"Ascoli","year":"1982","journal-title":"J Biol Chem"},{"key":"2020071613564708800_R49","doi-asserted-by":"crossref","first-page":"2098","DOI":"10.1210\/endo.133.5.8404659","article-title":"A polyclonal antibody to a synthetic peptide derived from the rat FSH receptor reveals the recombinant receptor as a 74 kDa protein.","volume":"133","author":"Quintana","year":"1993","journal-title":"Endocrinology"},{"key":"2020071613564708800_R50","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1083\/jcb.145.1.123","article-title":"The receptor recycling pathway contains two distinct populations of early endosomes with different sorting functions.","volume":"145","author":"Sheff","year":"1999","journal-title":"J Cell Biol"},{"key":"2020071613564708800_R51","first-page":"906","article-title":"Panning transfected cells for electrophysiological studies.","volume":"15","author":"Margolskee","year":"1993","journal-title":"Biotechniques"},{"key":"2020071613564708800_R52","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1073\/pnas.75.1.99","article-title":"Gonadotropin binding and stimulation of steroidogenesis in Leydig tumor cells.","volume":"75","author":"Ascoli","year":"1978","journal-title":"Proc Natl Acad Sci USA"}],"container-title":["Molecular Endocrinology"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/mend\/article-pdf\/20\/11\/2931\/10720417\/mend2931.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/mend\/article-pdf\/20\/11\/2931\/10720417\/mend2931.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T16:07:56Z","timestamp":1736438876000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/mend\/article\/20\/11\/2931\/2738592"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,11,1]]},"references-count":52,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2006,11,1]]}},"URL":"https:\/\/doi.org\/10.1210\/me.2006-0138","relation":{},"ISSN":["0888-8809","1944-9917"],"issn-type":[{"type":"print","value":"0888-8809"},{"type":"electronic","value":"1944-9917"}],"subject":[],"published":{"date-parts":[[2006,11,1]]}}}