Engineering bromodomains with a photoactive amino acid by engaging 'Privileged' tRNA synthetases

• Published in: Chemical communications (Cambridge, England) Vol. 56; no. 25; pp. 3641 - 3644
• Main Authors: Wagner, Shana, Sudhamalla, Babu, Mannes, Philip, Sappa, Sushma, Kavoosi, Sam, Dey, Debasis, Wang, Sinan, Islam, Kabirul
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 28.03.2020
• Subjects: Amino acids; Amino Acids - chemistry; Amino Acids - metabolism; Amino Acyl-tRNA Synthetases - chemistry; Amino Acyl-tRNA Synthetases - metabolism; Cross-Linking Reagents - chemical synthesis; Cross-Linking Reagents - chemistry; Cross-Linking Reagents - metabolism; Crosslinking; Crystallography; Molecular Structure; Phenylalanine; Phenylalanine - analogs & derivatives; Phenylalanine - chemistry; Phenylalanine - metabolism; Photochemical Processes; Protein Engineering
• ISSN: 1359-7345; 1364-548X
• DOI: 10.1039/c9cc09891g

Site-specific placement of unnatural amino acids, particularly those responsive to light, offers an elegant approach to control protein function and capture their fleeting 'interactome'. Herein, we have resurrected 4-(trifluoromethyldiazirinyl)-phenylalanine, an underutilized photo-crosslinker, by introducing several key features including easy synthetic access, site-specific incorporation by 'privileged' synthetases and superior crosslinking efficiency, to develop photo-crosslinkable bromodomains suitable for 'interactome' profiling. An improved chemical synthesis, site-specific incorporation and enhanced photo-crosslinking ability of tmdF have been demonstrated in the context of protein-protein interactions.