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

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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Pirman NL,Barber KW,Aerni HR,Ma NJ,Haimovich AD,Rogulina S,Isaacs FJ,Rinehart J

doi

10.1038/ncomms9130

subject

Has Abstract

pub_date

2015-09-09 00:00:00

pages

8130

issn

2041-1723

pii

ncomms9130

journal_volume

6

pub_type

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