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Optimal Conditions for Production of B Class G Protein-Coupled Receptor Expressed from E. coli

초록

영어

G protein-coupled-receptors (GPCRs) are a family of integral membrane proteins with seven transmembrane helices involved in a range of physiological processes, including cell signal transduction, sensory signaling, neuronal transmission and hormonal signaling. GPCRs are associated with almost every major therapeutic category or disease class, including pain,
asthma, inflammation, obesity and cancer. More than 50% of all drugs target GPCRs and control their activities. Although many efforts have been given to use E. coli as host cell for the production of GPCRs, it is difficult to express because of their strong hydrophobicity and complicated structure. In this study, human parathyroid hormone receptor type 1, B class GPCR,
was expressed in E. coli at high-level mainly as insoluble form after cloning of the gene into bacterial expressing vector, pDEST15. The expressed receptor was successfully solubilized with anionic detergent for further purification. This would be the first example for B class GPCR produced form E. coli in miligram quantity which is suitable amount for high-resolution structural
techniques, such as solid-state NMR and crystallography. This B class GPCR production method can be applied to development of GPCR biosensor that might be wildly used in the field of diagnosis and drug discovery.

저자정보

  • Sae Ryun AHN School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742.
  • Hyun Seok SONG School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742.
  • Sang Hun LEE School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742.
  • Eun Hae OH School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742.
  • Tai Hyun PARK School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742.

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