2YXN

Structual basis of azido-tyrosine recognition by engineered bacterial Tyrosyl-tRNA synthetase

Summary for 2YXN

• Entry DOI: 10.2210/pdb2yxn/pdb
• Related: 1WQ3
• Descriptor: Tyrosyl-tRNA synthetase, 3-AZIDO-L-TYROSINE (3 entities in total)
• Functional Keywords: trna synthetases class i, structural genomics, nppsfa, national project on protein structural and functional analyses, riken structural genomics/proteomics initiative, rsgi, ligase
• Biological source: Escherichia coli
• Total number of polymer chains: 1
• Total formula weight: 36206.95

Authors

Oki, K.,Kobayashi, T.,Sakamoto, K.,Yokoyama, S.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (deposition date: 2007-04-26, release date: 2008-04-29, Last modification date: 2023-10-25)

Primary citation

Iraha, F.,Oki, K.,Kobayashi, T.,Ohno, S.,Yokogawa, T.,Nishikawa, K.,Yokoyama, S.,Sakamoto, K.
Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRNATyr pair by the archaeal tyrosine pair in Escherichia coli for genetic code expansion
Nucleic Acids Res., 38:3682-3691, 2010

PubMed Abstract: Non-natural amino acids have been genetically encoded in living cells, using aminoacyl-tRNA synthetase-tRNA pairs orthogonal to the host translation system. In the present study, we engineered Escherichia coli cells with a translation system orthogonal to the E. coli tyrosyl-tRNA synthetase (TyrRS)-tRNA(Tyr) pair, to use E. coli TyrRS variants for non-natural amino acids in the cells without interfering with tyrosine incorporation. We showed that the E. coli TyrRS-tRNA(Tyr) pair can be functionally replaced by the Methanocaldococcus jannaschii and Saccharomyces cerevisiae tyrosine pairs, which do not cross-react with E. coli TyrRS or tRNA(Tyr). The endogenous TyrRS and tRNA(Tyr) genes were then removed from the chromosome of the E. coli cells expressing the archaeal TyrRS-tRNA(Tyr) pair. In this engineered strain, 3-iodo-L-tyrosine and 3-azido-L-tyrosine were each successfully encoded with the amber codon, using the E. coli amber suppressor tRNATyr and a TyrRS variant, which was previously developed for 3-iodo-L-tyrosine and was also found to recognize 3-azido-L-tyrosine. The structural basis for the 3-azido-L-tyrosine recognition was revealed by X-ray crystallography. The present engineering allows E. coli TyrRS variants for non-natural amino acids to be developed in E. coli, for use in both eukaryotic and bacterial cells for genetic code expansion.

PubMed: 20159998
DOI: 10.1093/nar/gkq080

Experimental method

X-RAY DIFFRACTION (1.8 Å)