Glycosylation of the calcitonin receptor-like receptor at Asn(60) or Asn(112) is important for cell surface expression.
The 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 Asn(60), Asn(112) and Asn(117) 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 [(125)I]CGRP and -ADM binding. Similarly, the double mutant myc-hCRLR(N60,112T) exhibited minimal N-glycosidase F sensitive glycosylation, presumably at the third Asn(117), and the cell surface expression and specific radioligand binding were impaired. Substitution of the Asn(117) by Thr abolished CGRP and ADM binding in the face of intact N-glycosylation and cell surface expression.
Bühlmann
N
N
Research Laboratory for Calcium Metabolism, Department of Orthopaedic Surgery, University of Zurich, Klinik Balgrist, Forchstrasse 340, 8008, Zurich, Switzerland.
Aldecoa
A
A
Leuthäuser
K
K
Gujer
R
R
Muff
R
R
Fischer
J A
JA
Born
W
W
D053607:Adrenomedullin; D000581:Amidohydrolases / Q000378:metabolism; D001216:Asparagine / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000187:drug effects / Q000235:genetics; D001667:Binding, Competitive / Q000187:drug effects / Q000235:genetics; D015153:Blotting, Western; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D058286:Calcitonin Receptor-Like Protein; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D016297:Mutagenesis, Site-Directed; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010455:Peptides / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D011869:Radioligand Assay; D018003:Receptors, Calcitonin / Q000235:genetics / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology
C545578:CALCRL protein, human; D058286:Calcitonin Receptor-Like Protein; D010455:Peptides; D016271:Proto-Oncogene Proteins c-myc; D018003:Receptors, Calcitonin; D014415:Tunicamycin; D053607:Adrenomedullin; D001216:Asparagine; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D015740:Calcitonin Gene-Related Peptide