通过邻近增强生物反应在蛋白质中添加非天然共价键。
Adding an unnatural covalent bond to proteins through proximity-enhanced bioreactivity.
机构信息
The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, La Jolla, California, USA.
出版信息
Nat Methods. 2013 Sep;10(9):885-8. doi: 10.1038/nmeth.2595. Epub 2013 Aug 4.
Abstract
Natural proteins often rely on the disulfide bond to covalently link side chains. Here we genetically introduce a new type of covalent bond into proteins by enabling an unnatural amino acid to react with a proximal cysteine. We demonstrate the utility of this bond for enabling irreversible binding between an affibody and its protein substrate, capturing peptide-protein interactions in mammalian cells, and improving the photon output of fluorescent proteins.
摘要
天然蛋白质通常依赖于二硫键将侧链共价连接。在这里，我们通过使非天然氨基酸与邻近的半胱氨酸反应，在蛋白质中遗传引入一种新的共价键。我们证明了这种键在使亲和体与其蛋白质底物之间形成不可逆结合、捕获哺乳动物细胞中的肽-蛋白质相互作用以及提高荧光蛋白的光子输出方面的用途。
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