title: Identification of N-glycosylation sites in the gonadotropin-releasing hormone receptor: role in receptor expression but not ligand binding.
abstract: The asparagine residues of the three N-glycosylation consensus sequences in the mouse gonadotropin-releasing hormone receptor were mutated to determine which residues were glycosylated and the function of glycosylation. Photoaffinity labelled Gln4 and Gln18 receptor mutants exhibited lower apparent molecular weight on SDS polyacrylamide gel electrophoresis, while the Gln102 receptor showed wildtype mobility. This indicates that the receptor is glycosylated at Asn4 and Asn18 but not at Asn102. Binding affinities of all the mutant receptors were normal, indicating that carbohydrate moieties are not involved in ligand binding interactions. However, expression of the Gln4 and Gln18 receptors were substantially decreased, indicating a role for glycosylation in receptor expression or stability. All the glycosylation site mutants were capable of normal signal transduction, as indicated by their ability to stimulate inositol phosphate production.
authors.affiliation: MRC Regulatory Peptides Research Unit, University of Cape Town Medical School, South Africa.
authors.forename: J S
authors.initials: JS
authors.lastname: Davidson
authors.forename: C A
authors.initials: CA
authors.lastname: Flanagan
authors.forename: W
authors.initials: W
authors.lastname: Zhou
authors.forename: I I
authors.initials: II
authors.lastname: Becker
authors.forename: R
authors.initials: R
authors.lastname: Elario
authors.forename: W
authors.initials: W
authors.lastname: Emeran
authors.forename: S C
authors.initials: SC
authors.lastname: Sealfon
authors.forename: R P
authors.initials: RP
authors.lastname: Millar
mesh_terms: D000818:Animals; D001216:Asparagine / Q000737:chemistry; D001665:Binding Sites; D002460:Cell Line; D016384:Consensus Sequence; D006031:Glycosylation; D007295:Inositol Phosphates / Q000096:biosynthesis; D008024:Ligands; D051379:Mice; D016297:Mutagenesis, Site-Directed; D011966:Receptors, LHRH / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D007295:Inositol Phosphates; D008024:Ligands; D011966:Receptors, LHRH; D001216:Asparagine