Genetically Encoding Fluorosulfate- l -tyrosine To React with Lysine, Histidine, and Tyrosine via SuFEx in Proteins in Vivo

作者：Nanxi Wang, Bing Yang, Caiyun Fu, He Zhu, Feng Zheng, Tomonori Kobayashi, Jun Liu, Shanshan Li, Cheng Ma, Peng George Wang, Qian Wang, Lei Wang · 发表于：Journal of the American Chemical Society · 年份：2018 · DOI：10.1021/jacs.8b01087 · 被引用次数：246 · 研究领域：Click Chemistry and Applications、Chemical Synthesis and Analysis、Monoclonal and Polyclonal Antibodies Research

Introducing new chemical reactivity into proteins in living cells would endow innovative covalent bonding ability to proteins for research and engineering in vivo. Latent bioreactive unnatural amino acids (Uaas) can be incorporated into proteins to react with target natural amino acid residues via proximity-enabled reactivity. To expand the diversity of proteins amenable to such reactivity in vivo, a chemical functionality that is biocompatible and able to react with multiple natural residues under physiological conditions is highly desirable. Here we report the genetic encoding of fluorosulfate-l-tyrosine (FSY), the first latent bioreactive Uaa that undergoes sulfur-fluoride exchange (SuFEx) on proteins in vivo. FSY was found nontoxic to Escherichia coli and mammalian cells; after being incorporated into proteins, it selectively reacted with proximal lysine, histidine, and tyrosine via SuFEx, generating covalent intraprotein bridge and interprotein cross-link of interacting proteins directly in living cells. The proximity-activatable reactivity, multitargeting ability, and excellent biocompatibility of FSY will be invaluable for covalent manipulation of proteins in vivo. Moreover, genetically encoded FSY hereby empowers general proteins with the next generation of click chemistry, SuFEx, which will afford broad utilities in chemical biology, drug discovery, and biotherapeutics.