Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology, Tokyo, Japan.
J Biomol NMR. 2010 Jan;46(1):3-10. doi: 10.1007/s10858-009-9377-0. Epub 2009 Sep 29.
The preparation of stable isotope-labeled proteins is necessary for the application of a wide variety of NMR methods, to study the structures and dynamics of proteins and protein complexes. The E. coli expression system is generally used for the production of isotope-labeled proteins, because of the advantages of ease of handling, rapid growth, high-level protein production, and low cost for isotope-labeling. However, many eukaryotic proteins are not functionally expressed in E. coli, due to problems related to disulfide bond formation, post-translational modifications, and folding. In such cases, other expression systems are required for producing proteins for biomolecular NMR analyses. In this paper, we review the recent advances in expression systems for isotopically labeled heterologous proteins, utilizing non-E. coli prokaryotic and eukaryotic cells.
稳定同位素标记蛋白的制备对于广泛应用各种 NMR 方法研究蛋白质和蛋白质复合物的结构和动力学是必要的。由于操作简便、生长迅速、蛋白质产量高、同位素标记成本低等优点，大肠杆菌表达系统通常用于同位素标记蛋白的生产。然而，由于涉及二硫键形成、翻译后修饰和折叠等问题，许多真核蛋白在大肠杆菌中没有功能表达。在这种情况下，需要使用其他表达系统来生产用于生物分子 NMR 分析的蛋白质。本文综述了利用非大肠杆菌原核和真核细胞表达异源蛋白的同位素标记表达系统的最新进展。
