Internalization and recycling of receptor-bound gonadotropin-releasing hormone agonist in pituitary gonadotropes.
The fate of cell surface gonadotropin-releasing hormone (GnRH) receptors on pituitary cells was studied utilizing lysosomotropic agents and monensin. Labeling of pituitary cells with a photoreactive GnRH derivative, [azidobenzoyl-D-Lys6]GnRH, revealed a specific band of Mr = 60,000. When photoaffinity-labeled cells were exposed to trypsin immediately after completion of the binding, the radioactivity incorporated into the Mr = 60,000 band decreased, with a concomitant appearance of a proteolytic fragment (Mr = 45,000). This fragment reflects cell surface receptors. Following GnRH binding, the hormone-receptor complexes underwent internalization, partial degradation, and recycling. The process of hormone-receptor complex degradation was substantially prevented by lysosomotropic agents, such as chloroquine and methylamine, or the proton ionophore, monensin. Chloroquine and monensin, however, did not affect receptor recycling, since the tryptic fragment of Mr = 45,000 was evident after treatment with these agents. This suggests that recycling of GnRH receptors in gonadotropes occurs whether or not the internal environment is acidic. Based on these findings, we propose a model describing the intracellular pathway of GnRH receptors.
Schvartz
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Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
Hazum
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D000345:Affinity Labels / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000378:metabolism*; D002064:Buserelin / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000378:metabolism; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology; D010777:Photochemistry; D010902:Pituitary Gland / Q000378:metabolism*; D011966:Receptors, LHRH / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D007457:Iodine Radioisotopes; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone; C031737:LHRH, N-epsilon-azidobenzoyl-Lys(6)-; D002738:Chloroquine; D008985:Monensin; D014357:Trypsin; D002064:Buserelin