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- Title
Characterization of Cell Lines Stably Expressing Human Normal or Mutated EGFP-Tagged MC4R.
- Authors
Blondet, Antonine; Doghman, Mabrouka; Rached, Mohamed; Durand, Philippe; Bégeot, Martine; Naville, Danielle
- Abstract
The melanocortin receptor type 4 (MC4-R) is involved in food intake and represents a potential target for the treatment of some forms of obesity. The fluorescent protein EGFP was fused to the wild-type or mutated coding sequence of the human MC4-R. After transfection in HEK 293, clones stably expressing hMC4-R-EGFP were selected. Wild-type chimeric hMC4-R was well addressed to the cell membrane as demonstrated using confocal microscopy and displayed the same pharmacological characteristics as native hMC4R. NDP-αMSH induced a time-dependent internalization of MC4-R that was partially prevented by AgRP. The two mutated chimeric receptors studied here (CTCT-deleted and C271A) showed a high alteration of their response to ligand and were retained inside the cells. In conclusion, we have developed a model of clones stably expressing EGFP-tagged-hMC4-R. This is the only such model available to date and it provides a useful tool to follow the trafficking of MC4-R inside living cells.
- Subjects
CELL lines; CELL culture; MSH (Hormone); GENETIC transformation; VIRAL genetics; NUTRITION disorders
- Publication
Journal of Biochemistry, 2004, Vol 135, Issue 4, p541
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvh064