Genetically encoded protein photocrosslinker with a transferable mass spectrometry-identifiable label
Citations Over TimeTop 10% of 2016 papers
Abstract
Coupling photocrosslinking reagents with mass spectrometry has become a powerful tool for studying protein-protein interactions in living systems, but it still suffers from high rates of false-positive identifications as well as the lack of information on interaction interface due to the challenges in deciphering crosslinking peptides. Here we develop a genetically encoded photo-affinity unnatural amino acid that introduces a mass spectrometry-identifiable label (MS-label) to the captured prey proteins after photocrosslinking and prey-bait separation. This strategy, termed IMAPP (In-situ cleavage and MS-label transfer After Protein Photocrosslinking), enables direct identification of photo-captured substrate peptides that are difficult to uncover by conventional genetically encoded photocrosslinkers. Taking advantage of the MS-label, the IMAPP strategy significantly enhances the confidence for identifying protein-protein interactions and enables simultaneous mapping of the binding interface under living conditions.
Related Papers
- → Improved efficiency of site-specific copper(II) ion-catalysed protein cleavage effected by mutagenesis of cleavage site(2000)28 cited
- → Factor Xa cleavage of fusion proteins(1990)30 cited
- → Design and Applications of Genetically Engineered Nanocomposites(2010)1 cited
- → Cavity formation at cleavage cracks in KBr:Sn(1965)
- → Cleavage(1960)