Italia James S, Zheng Yunan, Kelemen Rachel E, Erickson Sarah B, Addy Partha S, Chatterjee Abhishek
Department of Chemistry, Boston College, Chestnut Hill, MA, U.S.A.
Biochem Soc Trans. 2017 Apr 15;45(2):555-562. doi: 10.1042/BST20160336.
In the last two decades, unnatural amino acid (UAA) mutagenesis has emerged as a powerful new method to probe and engineer protein structure and function. This technology enables precise incorporation of a rapidly expanding repertoire of UAAs into predefined sites of a target protein expressed in living cells. Owing to the small footprint of these genetically encoded UAAs and the large variety of enabling functionalities they offer, this technology has tremendous potential for deciphering the delicate and complex biology of the mammalian cells. Over the last few years, exciting progress has been made toward expanding the toolbox of genetically encoded UAAs in mammalian cells, improving the efficiency of their incorporation and developing innovative applications. Here, we provide our perspective on these recent developments and highlight the current challenges that must be overcome to realize the full potential of this technology.
在过去二十年中，非天然氨基酸（UAA）诱变已成为一种强大的新方法，用于探究和改造蛋白质的结构与功能。这项技术能够将迅速扩充的非天然氨基酸库精确地掺入活细胞中表达的目标蛋白质的预定义位点。由于这些基因编码的非天然氨基酸占用空间小且具有多种多样的功能特性，该技术在解读哺乳动物细胞微妙而复杂的生物学特性方面具有巨大潜力。在过去几年里，在扩充哺乳动物细胞中基因编码非天然氨基酸的工具库、提高其掺入效率以及开发创新应用方面取得了令人兴奋的进展。在此，我们阐述对这些最新进展的看法，并强调要充分发挥该技术的潜力必须克服的当前挑战。
