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

FEBS Letters

Born, Walter¹  Gujer, Remo¹  Bühlmann, Nicole¹  Fischer, Jan A¹  Muff, Roman¹  Aldecoa, Amaya¹  Leuthäuser, Kerstin¹ 

[1] Research Laboratory for Calcium Metabolism, Departments of Orthopaedic Surgery and Medicine, University of Zurich, Klinik Balgrist, Forchstrasse 340, 8008 Zurich, Switzerland

关键词: Adrenomedullin;    Calcitonin gene-related peptide;    Calcitonin receptor-like receptor;    Glycosylation;    Receptor-activity-modifying protein;    Tunicamycin;    ADM;    adrenomedullin;    CGRP;    calcitonin gene-related peptide;    CRLR;    calcitonin receptor-like receptor;    CT;    calcitonin;    CTR;    calcitonin receptor;    PTH;    parathyroid hormone;    PTHrP;    parathyroid hormone-related peptide;    RAMP;    human receptor-activity-modifying protein;    TSA;    SV40 T-antigen transformed human embryonic kidney cells;    VIP;    vasoactive intestinal polypeptide;   

DOI  :  10.1016/S0014-5793(00)02259-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 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.

【 授权许可】

Unknown