Wang Lei, Xie Jianming, Schultz Peter G
The Jack H. Skirball Center for Chemical Biology & Proteomics, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
Annu Rev Biophys Biomol Struct. 2006;35:225-49. doi: 10.1146/annurev.biophys.35.101105.121507.
Recently, a general method was developed that makes it possible to genetically encode unnatural amino acids with diverse physical, chemical, or biological properties in Escherichia coli, yeast, and mammalian cells. More than 30 unnatural amino acids have been incorporated into proteins with high fidelity and efficiency by means of a unique codon and corresponding tRNA/aminoacyl-tRNA synthetase pair. These include fluorescent, glycosylated, metal-ion-binding, and redox-active amino acids, as well as amino acids with unique chemical and photochemical reactivity. This methodology provides a powerful tool both for exploring protein structure and function in vitro and in vivo and for generating proteins with new or enhanced properties.
最近,人们开发了一种通用方法,该方法能够在大肠杆菌、酵母和哺乳动物细胞中对具有多种物理、化学或生物学特性的非天然氨基酸进行基因编码。借助独特的密码子以及相应的tRNA/氨酰-tRNA合成酶对,已有30多种非天然氨基酸以高保真度和高效率掺入蛋白质中。这些氨基酸包括荧光、糖基化、金属离子结合和氧化还原活性氨基酸,以及具有独特化学和光化学反应性的氨基酸。这种方法为在体外和体内探索蛋白质结构和功能以及生成具有新特性或增强特性的蛋白质提供了一个强大的工具。