Phosphorylation and glycosylation of the luteinizing hormone receptor.
Purified testicular and ovarian luteinizing hormone/human chorionic gonadotropin (hCG) receptors are phosphorylated at serine and threonine residues by the catalytic subunit of the cAMP-dependent protein kinase (protein kinase A). Occupancy of the receptors by hCG significantly increased the rate but not the extent of phosphorylation. However, prolonged preincubation of receptors with hCG reduced the subsequent rate of receptor phosphorylation. Identical phosphopeptide maps were obtained for the phosphorylated ovarian and testicular receptors. The phosphorylated receptor, like the native receptor, bound to wheat germ lectin and hCG-Sepharose and migrated as a single band of Mr 90,000 (testis) and Mr 85,000 (ovary) on NaDodSO4/PAGE. Neuraminidase treatment of receptors caused reductions of molecular weight to 82,000 (testis) and 77,000 (ovary), and further treatment with O-Glycanase had minimal effect on molecular size. However, deglycosylation with N-Glycosidase and endoglycosidase F produced a single labeled polypeptide of Mr 59,000 for both gonadal receptors. Treatment of native receptors with neuraminidase caused no apparent change in binding of gonadotropin to blotted receptors, whereas deglycosylated receptors showed a major reduction in hormone binding. These results indicate that luteinizing hormone/hCG receptors are sialoglycoproteins with predominantly N-linked glycosyl residues that account for the size difference between testicular and ovarian receptors and that may participate in the interaction with gonadotropin. Receptor occupancy by agonist leads to a conformational change that facilitates its phosphorylation during initial binding and reduces the rate of phosphorylation after more prolonged exposure to hCG.
Minegishi
T
T
Molecular and Endocrinology Section, National Institute of Child Health and Human Development, Bethesda, MD 20892.
Delgado
C
C
Dufau
M L
ML
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005260:Female; D006031:Glycosylation; D007700:Kinetics; D008297:Male; D010053:Ovary / Q000378:metabolism; D010449:Peptide Mapping; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D011974:Receptors, LH / Q000187:drug effects / Q000378:metabolism*; D013737:Testis / Q000378:metabolism
D000596:Amino Acids; D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D011494:Protein Kinases