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Expanding the genetic code of Escherichia coli with phosphoserine.

ABSTRACT: O-Phosphoserine (Sep), the most abundant phosphoamino acid in the eukaryotic phosphoproteome, is not encoded in the genetic code, but synthesized posttranslationally. Here, we present an engineered system for specific cotranslational Sep incorporation (directed by UAG) into any desired position in a protein by an Escherichia coli strain that harbors a Sep-accepting transfer RNA (tRNA(Sep)), its cognate Sep-tRNA synthetase (SepRS), and an engineered EF-Tu (EF-Sep). Expanding the genetic code rested on reengineering EF-Tu to relax its quality-control function and permit Sep-tRNA(Sep) binding. To test our system, we synthesized the activated form of human mitogen-activated ERK activating kinase 1 (MEK1) with either one or two Sep residues cotranslationally inserted in their canonical positions (Sep(218), Sep(222)). This system has general utility in protein engineering, molecular biology, and disease research.

SUBMITTER: Park HS

PROVIDER: S-EPMC5547737 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature

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Expanding the genetic code of Escherichia coli with phosphoserine.

Park Hee-Sung HS Hohn Michael J MJ Umehara Takuya T Guo Li-Tao LT Osborne Edith M EM Benner Jack J Noren Christopher J CJ Rinehart Jesse J Söll Dieter D

Science (New York, N.Y.) 20110801 6046

O-Phosphoserine (Sep), the most abundant phosphoamino acid in the eukaryotic phosphoproteome, is not encoded in the genetic code, but synthesized posttranslationally. Here, we present an engineered system for specific cotranslational Sep incorporation (directed by UAG) into any desired position in a protein by an Escherichia coli strain that harbors a Sep-accepting transfer RNA (tRNA(Sep)), its cognate Sep-tRNA synthetase (SepRS), and an engineered EF-Tu (EF-Sep). Expanding the genetic code rest ...[more]

PMID: 21868676

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