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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.

SUBMITTER: Pirman NL

PROVIDER: S-EPMC4566969 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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

Pirman Natasha L NL Barber Karl W KW Aerni Hans R HR Ma Natalie J NJ Haimovich Adrian D AD Rogulina Svetlana S Isaacs Farren J FJ Rinehart Jesse J

Nature communications 20150909

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 demo ...[more]

PMID: 26350500

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