Expanding the genetic code of Escherichia coli with phosphotyrosine
by Fan, Chenguang; Ip, Kevan; Soell, Dieter
Protein phosphorylation is one of the most important post-translational modifications in nature. However, the site-specific incorporation of O-phosphotyrosine into proteins in vivo has not yet been reported. Endogenous phosphatases present in cells can dephosphorylate phosphotyrosine as a free amino acid or as a protein residue. Therefore, we deleted the genes of five phosphatases from the genome of Escherichia coli with the aim of stabilizing phosphotyrosine. Together with an engineered aminoacyl-tRNA synthetase (derived from Methanocaldococcus jannaschii tyrosyl-tRNA synthetase) and an elongation factor Tu variant, we were able to cotranslationally incorporate O-phosphotyrosine into the superfolder green fluorescent protein at a desired position in vivo. This system will facilitate future studies of tyrosine phosphorylation.
- Journal
- FEBS Letters
- Volume
- 590
- Issue
- 17
- Year
- 2016
- Start Page
- 3040-3047
- URL
- https://dx.doi.org/10.1002/1873-3468.12333
- ISBN/ISSN
- 1873-3468; 0014-5793
- DOI
- 10.1002/1873-3468.12333