Chemical engineering of bacterial effectors for regulating cell signaling and responses

Capture and mass spectrometry analysis of effector-substrate complexes using genetically incorporated photo-crosslinkers in host cells

“…Uaas can be inserted into specific positions of proteins by engineered aminoacyl-tRNA synthetase and tRNA CUA pairs. The engineered aminoacyl-tRNA synthetase specifically recognizes and charges the Uaa onto the orthogonal tRNA CUA , which recognizes the antisense codon TAG to incorporate the Uaa into corresponding sites of protein during translation 6 , 7 , 8 , 9 , 10 , 11 , 12 ( Figure 1 B). When the photo-crosslinkers replaced the residues on the interaction surface of protein complexes, they react with adjacent amino acids to form covalent bond under ultraviolet treatment.…”