Randomized trial demonstrates enhanced protein labeling using H-L-photo-lysine for studying SIRT6 interactions.
Key Points
The research aims to develop a method for synthesizing a novel noncanonical amino acid, H-L-photo-lysine, and demonstrate its incorporation into proteins for studying interactions.
Synthesis of H-L-photo-lysine with improved yields and simplified purification
Genetic code expansion to incorporate photoK into a GFP variant
Photocrosslinking studies conducted with SIRT6K15photoK mutant in HEK293 and E. coli cell lysates
Successfully synthesized H-L-photo-lysine with improved yield and milder conditions
Confirmed UV-dependent crosslinking of SIRT6K15photoK with histone H3, indicating specific interactions
Pretreatment with a SIRT6-selective inhibitor significantly reduced crosslinking, confirming specificity of interactions