원문정보
초록
영어
Olfactory receptors, G protein-coupled receptor (GPCR), are integral membrane proteins composed of seven transmembrane spanning domains. These are not easy to be overexpressed, solubilized and purified because of their structures and strong hydrophobicity. Purification of olfactory receptor, where only a few studies have been done, is critical step for developing protein-based olfactory biosensor. In this work, human olfactory receptor, OR2AG1, was overexpressed in Escherichia coli. Electrophoresis of the overexpressed protein, visualized with Coomassie Blue staining and by immunoblotting with glutathione S-transferase (GST) antibody, revealed bands of 53, 118 and 160KDa, which were identified as the monomeric, dimeric and trimeric forms of the receptor proteins, respectively. The protein was difficult to solubilize with many detergents and only N-lauroyl sarcosine was found to be suitable for efficient solubilization. Human olfactory receptor, OR2AG1, was purified and solubilized by N-Lauroyl Sarcosine after treating membrane fraction with Triton X-100. Purified protein can be used as the primary transducer of olfactory biosensor after reconstitution in phospholipid vesicles.