Characterization of cell lines stably expressing human normal or mutated EGFP-tagged MC4R.

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-alpha MSH 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.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Blondet A,Doghman M,Rached M,Durand P,Bégeot M,Naville D

doi

10.1093/jb/mvh064

keywords:

subject

Has Abstract

pub_date

2004-04-01 00:00:00

pages

541-6

issue

4

eissn

0021-924X

issn

1756-2651

journal_volume

135

pub_type

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