Preparing your results

Our searching services are busy right now. Your search will reload in five seconds.

X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

Expanding the genetic code of Escherichia coli with phosphoserine.

Science (New York, N.Y.) | Aug 26, 2011

http://www.ncbi.nlm.nih.gov/pubmed/21868676

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.

Pubmed ID: 21868676 RIS Download

Mesh terms: Amino Acyl-tRNA Synthetases | Anticodon | Chloramphenicol | Chloramphenicol O-Acetyltransferase | Codon, Terminator | Drug Resistance, Bacterial | Escherichia coli | Genetic Code | Genetic Engineering | Humans | MAP Kinase Kinase 1 | Peptide Elongation Factor Tu | Phosphoserine | Protein Engineering | Protein Modification, Translational | RNA, Bacterial | RNA, Transfer, Amino Acid-Specific | RNA, Transfer, Amino Acyl | RNA, Transfer, Cys | Recombinant Fusion Proteins | Transfer RNA Aminoacylation

Research resources used in this publication

None found

Research tools detected in this publication

None found

Data used in this publication

None found

Associated grants

  • Agency: NIGMS NIH HHS, Id: GM22854
  • Agency: NIDDK NIH HHS, Id: K01DK089006

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.