N-linked oligosaccharides are not required for hormone binding of the lutropin receptor in a Leydig tumor cell line and rat granulosa cells.
We have examined roles of carbohydrates of the lutropin receptor in a murine Leydig tumor cell line (MLTC) and primary cultures of rat granulosa cells. We approached this issue by deglycosylating mature receptors with glycosidases and by preventing glycosylation of nascent receptors with tunicamycin B2, an inhibitor of protein glycosylation but not protein synthesis. Deglycosylation of mature receptors with neuraminidase, N-glycanase or both did not affect ligand binding capacity. Regardless of glycosidase treatment, the number of hormone binding sites was similar. The Kas for native receptors, asialoreceptors and aglycoreceptors, are also comparable, being 2.0 x 10(9) M-1, 1.9 x 10(9) M-1 and 1.7 x 10(9) M-1 respectively. In contrast, cells treated with tunicamycin B2 failed to bind the hormone. These results demonstrate that N-oligosaccharides of mature lutropin receptors are not required for ligand binding. In addition, our data suggest, for the first time, that N-glycosylation of the receptor may be necessary for expressing functional receptors on the cell surface and that there exist striking similarities in roles of oligosaccharides of lutropin and its receptor.
Ji
I H
IH
Department of Molecular Biology, University of Wyoming, Laramie 82071-3944.
Slaughter
R G
RG
Ji
T H
TH
D000581:Amidohydrolases / Q000494:pharmacology; D000818:Animals; D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000502:physiology*; D006063:Chorionic Gonadotropin / Q000378:metabolism; D005260:Female; D006031:Glycosylation; D006107:Granulosa Cells / Q000378:metabolism*; D007984:Leydig Cell Tumor / Q000378:metabolism*; D051379:Mice; D009439:Neuraminidase / Q000494:pharmacology; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D051381:Rats; D011974:Receptors, LH / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000031:analogs & derivatives / Q000494:pharmacology
D002241:Carbohydrates; D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D014415:Tunicamycin; C046159:tunicamycin B2; D009439:Neuraminidase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase