刊名:Biochemical and Biophysical Research Communications
出版年:2003
出版时间:June 20, 2003
年:2003
卷:306
期:1
页码:208-212
全文大小:142 K
文摘
Among the melanocortins α-MSH is known to be involved in feeding behavior. These hormones mediate their effects through G protein-coupled receptors by stimulating adenylate cyclase. In this study, we have developed an in vitro expression model for human melanocortin 3 receptor (hMC3R) tagged at its C terminus with EGFP. The corresponding chimeric cDNA was stably expressed in HEK293 cells. The selected clones expressing the hMC3R–EGFP exhibited cell surface fluorescence and responded to NDP-MSH stimulation by producing cAMP in a dose-dependent manner (EC50: 0.3nM). Binding studies revealed a single class of binding sites with a KD of 2.24nM. Moreover, Agouti-related protein was also demonstrated to be an antagonist of the hMC3R–EGFP. Thus, the hMC3R tagged with EGFP stably expressed in HEK293 cells, exhibiting the same characteristics than the wild-type hMC3R, is the only model of expression of this receptor allowing its direct localization inside living cells.
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