Addy Partha Sarathi, Erickson Sarah B, Italia James S, Chatterjee Abhishek
Department of Chemistry, Boston College, Chestnut Hill, MA, USA.
Methods Mol Biol. 2019;2033:239-251. doi: 10.1007/978-1-4939-9654-4_16.
Chemoselective protein labeling is a valuable tool in the arsenal of modern chemical biology. The unnatural amino acid mutagenesis technology provides a powerful way to site-specifically introduce nonnatural chemical functionalities into recombinant proteins, which can be subsequently functionalized in a chemoselective manner. Even though several strategies currently exist to selectively label recombinant proteins in this manner, there is considerable interest for the development of additional chemoselective reactions that are fast, catalyst-free, use readily available reagents, and are compatible with existing conjugation chemistries. Here we describe a method to express recombinant proteins in E. coli site-specifically incorporating 5-hydroxytryptophan, followed by the chemoselective labeling of this residue using a chemoselective rapid azo-coupling reaction.
化学选择性蛋白质标记是现代化学生物学手段中的一种重要工具。非天然氨基酸诱变技术提供了一种强大的方法，可将非天然化学官能团位点特异性地引入重组蛋白中，随后这些官能团可以通过化学选择性方式进行功能化修饰。尽管目前存在多种以这种方式选择性标记重组蛋白的策略，但人们仍对开发其他化学选择性反应有着浓厚兴趣，这些反应需具备快速、无催化剂、使用易得试剂且与现有偶联化学兼容的特点。在此，我们描述了一种在大肠杆菌中表达重组蛋白的方法，该方法可位点特异性地掺入5-羟色氨酸，随后利用化学选择性快速偶氮偶联反应对该残基进行化学选择性标记。
