Glycosylation of the calcitonin receptor-like receptor at Asn60 or Asn112 is important for cell surface expression

AbstractThe human calcitonin (CT) receptor-like receptor (hCRLR) of the B family of G protein-coupled receptors is N-glycosylated and associates with receptor-activity-modifying proteins for functional interaction with CT gene-related peptide (CGRP) or adrenomedullin (ADM), respectively. Three putative N-glycosylation sites Asn60, Asn112 and Asn117 are present in the amino-terminal extracellular domain of the hCRLR. Tunicamycin dose-dependently inhibited the glycosylation of a myc-tagged hCRLR and in parallel specific [125I]CGRP and -ADM binding. Similarly, the double mutant myc-hCRLR(N60,112T) exhibited minimal N-glycosidase F sensitive glycosylation, presumably at the third Asn117, and the cell surface expression and specific radioligand binding were impaired. Substitution of the Asn117 by Thr abolished CGRP and ADM binding in the face of intact N-glycosylation and cell surface expression