Design of an effective small expression tag to enhance GPCR production in E. coli-based cell-free and whole cell expression systems
- 주제(키워드) A/T-rich gene tag , cell-free expression , Escherichia coli , G protein-coupled receptor , protein expression , translation initiation rate
- 등재 SCIE, SCOPUS
- 발행기관 Wiley
- 발행년도 2023
- URI http://www.dcollection.net/handler/ewha/000000209990
- 본문언어 영어
- Published As https://doi.org/10.1002/pro.4839
- 저작권 이화여자대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
G protein-coupled receptors (GPCRs) play crucial roles in sensory, immune, and tumor metastasis processes, making them valuable targets for pharmacological and sensing applications in various industries. However, most GPCRs have low production yields in Escherichia coli (E. coli) expression systems. To overcome this limitation, we introduced AT10 tag, an effective fusion tag that could significantly enhance expression levels of various GPCRs in E. coli and its derived cell-free protein synthesis (CFPS) system. This AT10 tag consisted of an A/T-rich gene sequence designed via optimization of translation initiation rate. It is translated into a short peptide sequence of 10 amino acids at the N-terminus of GPCRs. Additionally, effector proteins could be utilized to suppress cytotoxicity caused by membrane protein expression, further boosting GPCR production in E. coli. Enhanced expression of various GPCRs using this AT10 tag is a promising approach for large-scale production of functional GPCRs in E. coli-based CFPS and whole cell systems, enabling their potential utilization across a wide range of industrial applications.
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