원문정보
초록
영어
G-protein coupled receptors (GPCRs), a family of integral membrane proteins with seven
transmembrane helices, are difficult to be overexpressed, solubilized and purified, especially from bacterial cell, because of their complicated structures and strong hydrophobicity. Before applying to chromatographic purification method, solubilization of GPCR expressed as inclusion body is critical step for GPCR production. In this work, a human olfactory receptor, hOR17-40, a family of GPCR, was overexpressed at high-level in E. coli mainly as insoluble form of inclusion body. Expressed human olfactory receptor was difficult to be solublilized by many detergents and chaotropes and sarcosyl and SDS was found to be suitable for effiient solubilization. Sarcosyl showed better solubilization ability than SDS and was thought to be the most suitable detergent for solubilization.
Solubilization of inclusion body was performed with different concentration of sarcosyl and
solubilized protein quantities were analyzed with immunoblotting and total protein assay. 5% sarcosyl was determined as optimal concentration for inclusion body solublization of the human olfactory receptor. After solubilization of GPCR, further chromatographic purification method can be applied to produce GPCR that is for being used in the fields of drug discovery and development of biosensor.