Addition of a photocrosslinking amino acid to the genetic code of Escherichia coli

作者:Chin JW; Martin AB; King DS; Wang L; Schultz PG*
来源:Proceedings of the National Academy of Sciences, 2002, 99(17): 11020-11024.
DOI:10.1073/pnas.172226299

摘要

Benzophenones are among the most useful photocrosslinking agents in biology. We have evolved an orthogonal aminoacyl-tRNA synthetase/tRNA pair that makes possible the in vivo incorporation of p-benzoyl-L-phenylalanine into proteins in Escherichia coli in response to the amber codon, TAG. This unnatural amino acid was incorporated with high translational efficiency and fidelity into the dimeric protein glutathione S-transferase. Irradiation resulted in efficient crosslinking (>50%) of the protein subunits. This methodology may prove useful for discovering and defining protein interactions in vitro and in vivo.

  • 出版日期2002-8-20