Nature Communications ( IF 16.6 ) Pub Date : 2020-09-24 , DOI: 10.1038/s41467-020-18629-9 Xinyuan He 1 , Yan Chen 2 , Daisy Guiza Beltran 3 , Maia Kelly 2 , Bin Ma 2 , Justin Lawrie 2 , Feng Wang 4 , Eric Dodds 2 , Limei Zhang 3 , Jiantao Guo 2 , Wei Niu 1
Protein tyrosine O-sulfation (PTS) plays a crucial role in extracellular biomolecular interactions that dictate various cellular processes. It also involves in the development of many human diseases. Regardless of recent progress, our current understanding of PTS is still in its infancy. To promote and facilitate relevant studies, a generally applicable method is needed to enable efficient expression of sulfoproteins with defined sulfation sites in live mammalian cells. Here we report the engineering, in vitro biochemical characterization, structural study, and in vivo functional verification of a tyrosyl-tRNA synthetase mutant for the genetic encoding of sulfotyrosine in mammalian cells. We further apply this chemical biology tool to cell-based studies on the role of a sulfation site in the activation of chemokine receptor CXCR4 by its ligand. Our work will not only facilitate cellular studies of PTS, but also paves the way for economical production of sulfated proteins as therapeutic agents in mammalian systems.
中文翻译:
哺乳动物细胞中磺基酪氨酸的功能性遗传编码
蛋白酪氨酸O硫酸化 (PTS) 在决定各种细胞过程的细胞外生物分子相互作用中起着至关重要的作用。它还涉及许多人类疾病的发展。不管最近的进展如何,我们目前对 PTS 的理解仍处于起步阶段。为了促进和促进相关研究,需要一种普遍适用的方法,以在活哺乳动物细胞中有效表达具有确定硫酸化位点的硫蛋白。在这里,我们报告了用于哺乳动物细胞中磺基酪氨酸遗传编码的酪氨酰-tRNA 合成酶突变体的工程、体外生化表征、结构研究和体内功能验证。我们进一步将此化学生物学工具应用于基于细胞的研究,研究硫酸化位点在其配体激活趋化因子受体 CXCR4 中的作用。