The development of a method to demonstrate the oligomerization of GPCRs

• Main Authors: Chuan Yi Yang, 楊川毅
• Other Authors: H. H. Lin
• Format: Others
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/08714069106885946866

碩士 === 長庚大學 === 生物醫學研究所 === 97 === The traditional method for the study of specific G protein-coupled receptors (GPCRs) interactions is to co-transfect two constructs at least in the beginning, but some limitations affecting the transfection efficiencies might cause unbalance of product amounts. For this reason, we attempted to improve the effects to develop a new manner for precise amounts of products. The internal ribosome entry site (IRES) bicistronic system was employed to determine the oligomerization of GPCRs. The EGF-like module containing mucin-like hormone receptor 2 (EMR2) and G protein-coupled receptor 56 (GPR56) receptors were listed in the same class of GPCR family. The protein products of IRES bicistronic constructs were analyzed for their interaction by co-immunoprecipitation (co-IP) and fluorescence-activated cell sorting manners (FACS). Our results indicated that the bicistronic system was with some defects at present, but we still verified further that the seven-transmembrane domain of ERM2 receptor is responsible for the formation of oligomers. In addition to EMR2 receptors, we considered that the GPR56 receptors do not interact with hetero-receptors.