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Nat Commun. 2015 Sep 9;6:8130. doi: 10.1038/ncomms9130.

A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation.

Author information

  • 1Department of Cellular &Molecular Physiology, Yale University, New Haven, Connecticut 06520-8114, USA.
  • 2Systems Biology Institute, Yale University, New Haven, Connecticut 06520-8114, USA.
  • 3Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8114, USA.

Abstract

Biochemical investigation of protein phosphorylation events is limited by inefficient production of the phosphorylated and non-phosphorylated forms of full-length proteins. Here using a genomically recoded strain of E. coli with a flexible UAG codon we produce site-specific serine- or phosphoserine-containing proteins, with purities approaching 90%, from a single recombinant DNA. Specifically, we synthesize human MEK1 kinase with two serines or two phosphoserines, from one DNA template, and demonstrate programmable kinase activity. Programmable protein phosphorylation is poised to help reveal the structural and functional information encoded in the phosphoproteome.

PMID:
26350500
PMCID:
PMC4566969
DOI:
10.1038/ncomms9130
[PubMed - indexed for MEDLINE]
Free PMC Article
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