A GFP fluorescence-based approach to determine detergent insolubility of the human serotonin1A receptor.
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 serotonin1A (5-HT1A) receptor. Our results show, for the first time, that a small yet significant fraction of the 5-HT1A 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-HT1A receptor in membrane domains and its significance in receptor function and signaling.
Kalipatnapu
Shanti
S
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India.
Chattopadhyay
Amitabha
A
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003902:Detergents; D017930:Genes, Reporter; D006801:Humans; D044282:Receptor, Serotonin, 5-HT1A / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D012995:Solubility; D014162:Transfection
D003902:Detergents; D011993:Recombinant Fusion Proteins; D044282:Receptor, Serotonin, 5-HT1A