Functional role of N‐linked glycosylation on the rat melanin‐concentrating hormone receptor 1

FEBS Letters

Kawamura, Yuuki¹  Tetsuka, Mitsue¹  Maruyama, Kei¹  Saito, Yumiko¹  Yue, Li¹ 

[1] Department of Pharmacology, Saitama Medical School, 38 Moro-Hongo, Moroyama-cho, Iruma-gun, Saitama 350-0495, Japan

关键词: Melanin-concentrating hormone;    G-protein-coupled receptor;    N-glycosylation site;    Calcium influx;    Ligand binding;    GPCR;    G-protein-coupled receptor;    MCH;    melanin-concentrating hormone;    MCHR1;    melanin-concentrating hormone receptor 1;    Flag-MCHR1;    amino-terminal Flag-tagged MCHR1;    FBS;    fetal bovine serum;    DMEM;    Dulbecco's modified Eagle's medium;    Endo H;    endoglycosidase H;    PNGase F;    peptide N-glycosidase F;    PBS;    phosphate-buffered saline;    SDS–PAGE;    sodium dodecyl sulfate–polyacrylamide gel electrophoresis;    FITC;    fluorescein isothiocyanate;   

DOI  :  10.1016/S0014-5793(02)03744-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Melanin-concentrating hormone (MCH) is known to act through two G-protein-coupled receptors MCHR1 and MCHR2. MCHR1 has three potential sites (Asn¹³, Asn¹⁶ and Asn²³) for N-linked glycosylation in its extracellular amino-terminus which may modulate its reactivity. Site-directed mutagenesis of the rat MCHR1 cDNA at single or multiple combinations of the three potential glycosylation sites was used to examine the role of the putative carbohydrate chains on receptor activity. It was found that all three potential N-linked glycosylation sites in MCHR1 were glycosylated, and that N-linked glycosylation of Asn²³ was necessary for full activity. Furthermore, disruption of all three glycosylation sites impaired proper expression at the cell surface and receptor activity. These data outline the importance of the N-linked glycosylation of the MCHR1.

【 授权许可】

Unknown