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

Nature Communications
Natasha L PirmanJesse Rinehart

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.

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Feb 26, 2016·Journal of Molecular Biology·Kevin C MaTimothy K Lu
Apr 21, 2016·FEBS Letters·Susanna GeorgePatrick O'Donoghue
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Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

SepOTSμ
Bio
SepOTS
Rad Image Lab

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