An Engineered Aryl Azide Ligase for Site‐Specific Mapping of Protein–Protein Interactions through Photo‐Cross‐Linking
作者： Hemanta Baruah , Sujiet Puthenveetil , Yoon-Aa Choi , Samit Shah , Alice Y. Ting
关键词: Binding site 、 Biochemistry 、 Azide 、 DNA ligase 、 Protein–protein interaction 、 FKBP 、 Protein engineering 、 Aryl 、 Fusion protein 、 Chemistry
摘要: We re-engineered the small-molecule binding site of E. coli lipoic acid ligase (LplA) to accept a fluorinated aryl azide probe in place acid. The mutated can covalently attach recombinant proteins fused 17-amino recognition sequence for LplA (called "LAP"). Labeling is highly specific, modifying LAP fusion exclusion all endogenous mammalian proteins. In cell lysate, we labeled FKBP with and demonstrated rapamycin-dependent photocrosslinking FRB.
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