R.F. School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853, USA.
R.F. School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853, USA.
Trends Biochem Sci. 2023 Jul;48(7):642-654. doi: 10.1016/j.tibs.2023.03.006. Epub 2023 Apr 20.
Despite advances in membrane protein (MP) structural biology and a growing interest in their applications, these proteins remain challenging to study. Progress has been hindered by the complex nature of MPs and innovative methods will be required to circumvent technical hurdles. Cell-free protein synthesis (CFPS) is a burgeoning technique for synthesizing MPs directly into a membrane environment using reconstituted components of the cellular transcription and translation machinery in vitro. We provide an overview of CFPS and how this technique can be applied to the synthesis and study of MPs. We highlight numerous strategies including synthesis methods and folding environments, each with advantages and limitations, to provide a survey of how CFPS techniques can advance the study of MPs.
尽管膜蛋白 (MP) 结构生物学取得了进展,并且人们对其应用越来越感兴趣,但这些蛋白质仍然难以研究。由于 MPs 的复杂性,研究进展受到阻碍,需要创新方法来规避技术障碍。无细胞蛋白质合成 (CFPS) 是一种新兴技术,可使用体外细胞转录和翻译机制的重组成分直接将 MPs 合成到膜环境中。我们提供了 CFPS 的概述,以及该技术如何应用于 MPs 的合成和研究。我们强调了许多策略,包括合成方法和折叠环境,每种方法都有其优点和局限性,以概述 CFPS 技术如何促进 MPs 的研究。