Site-specific incorporation of phosphotyrosine using an expanded genetic code.
- Author(s): Hoppmann, Christian
- Wong, Allison
- Yang, Bing
- Li, Shuwei
- Hunter, Tony
- Shokat, Kevan M
- Wang, Lei
- et al.
Published Web Locationhttps://doi.org/10.1038/nchembio.2406
Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phosphorylation in negative regulation of ubiquitination.