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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.
Abstract

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.

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