A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26350500.
- Also identified by DOI 10.1038/ncomms9130 and PMC identifier 4566969.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Codon, Terminator
- Escherichia coli
- Genome, Bacterial
- MAP Kinase Kinase 1
- Phosphorylation