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

作者：Jason W. Chin, Andrew Martin, DAVID SHAW KING, Lei Wang, Peter G. Schultz · 发表于：Proceedings of the National Academy of Sciences · 年份：2002 · DOI：10.1073/pnas.172226299 · 被引用次数：740 · 研究领域：RNA and protein synthesis mechanisms、RNA modifications and cancer、Bacterial Genetics and Biotechnology

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.