Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase

Tateki Suzuki, Corwin Miller, Li Tao Guo, Joanne M.L. Ho, David I. Bryson, Yane Shih Wang, David R. Liu, Dieter Söll

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Pyrrolysyl-tRNA synthetase (PylRS) is a major tool in genetic code expansion using noncanonical amino acids, yet its structure and function are not completely understood. Here we describe the crystal structure of the previously uncharacterized essential N-terminal domain of this unique enzyme in complex with tRNA Pyl . This structure explains why PylRS remains orthogonal in a broad range of organisms, from bacteria to humans. The structure also illustrates why tRNA Pyl recognition by PylRS is anticodon independent: the anticodon does not contact the enzyme. Then, using standard microbiological culture equipment, we established a new method for laboratory evolution - a noncontinuous counterpart of the previously developed phage-assisted continuous evolution. With this method, we evolved novel PylRS variants with enhanced activity and amino acid specificity. Finally, we employed an evolved PylRS variant to determine its N-terminal domain structure and show how its mutations improve PylRS activity in the genetic encoding of a noncanonical amino acid.

Original languageEnglish
Pages (from-to)1261-1266
Number of pages6
JournalNature Chemical Biology
Volume13
Issue number12
DOIs
Publication statusPublished - Dec 1 2017
Externally publishedYes

Fingerprint

Amino Acyl-tRNA Synthetases
Anticodon
Transfer RNA
Amino Acids
Genetic Code
Enzymes
Bacteriophages
Bacteria
Equipment and Supplies
Mutation

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Suzuki, T., Miller, C., Guo, L. T., Ho, J. M. L., Bryson, D. I., Wang, Y. S., ... Söll, D. (2017). Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase. Nature Chemical Biology, 13(12), 1261-1266. https://doi.org/10.1038/nchembio.2497

Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase. / Suzuki, Tateki; Miller, Corwin; Guo, Li Tao; Ho, Joanne M.L.; Bryson, David I.; Wang, Yane Shih; Liu, David R.; Söll, Dieter.

In: Nature Chemical Biology, Vol. 13, No. 12, 01.12.2017, p. 1261-1266.

Research output: Contribution to journalArticle

Suzuki, T, Miller, C, Guo, LT, Ho, JML, Bryson, DI, Wang, YS, Liu, DR & Söll, D 2017, 'Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase', Nature Chemical Biology, vol. 13, no. 12, pp. 1261-1266. https://doi.org/10.1038/nchembio.2497
Suzuki, Tateki ; Miller, Corwin ; Guo, Li Tao ; Ho, Joanne M.L. ; Bryson, David I. ; Wang, Yane Shih ; Liu, David R. ; Söll, Dieter. / Crystal structures reveal an elusive functional domain of pyrrolysyl-tRNA synthetase. In: Nature Chemical Biology. 2017 ; Vol. 13, No. 12. pp. 1261-1266.
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