A GFP fluorescence-based approach to determine detergent insolubility of the human serotonin_1A receptor

Abstract

Insolubility in non-ionic detergents such as Triton X-100 is a widely used biochemical criterion for characterization of membrane domains. We report here a novel green fluorescent protein fluorescence-based approach to directly determine detergent insolubility of specific membrane proteins. We have applied this method to explore the detergent resistance of an important G-protein coupled receptor, the serotonin_1A (5-HT_1A) receptor. Our results show, for the first time, that a small yet significant fraction of the 5-HT_1A receptor exhibits detergent insolubility. These results are validated by control experiments involving fluorescent lipid probes and protein markers. Our results assume relevance in the context of localization of the 5-HT_1A receptor in membrane domains and its significance in receptor function and signaling.