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Sequential Digestion with Trypsin and Elastase in Cross-Linking Mass Spectrometry.

Therese DauKapil GuptaImre BergerJuri Rappsilber
Published in: Analytical chemistry (2019)
Cross-linking mass spectrometry has become an important approach for studying protein structures and protein-protein interactions. The amino acid compositions of some protein regions impede the detection of cross-linked residues, although it would yield invaluable information for protein modeling. Here, we report on a sequential-digestion strategy with trypsin and elastase to penetrate regions with a low density of trypsin-cleavage sites. We exploited intrinsic substrate-recognition properties of elastase to specifically target larger tryptic peptides. Our application of this protocol to the TAF4-12 complex allowed us to identify cross-links in previously inaccessible regions.
Keyphrases
  • amino acid
  • mass spectrometry
  • high resolution
  • protein protein
  • liquid chromatography
  • binding protein
  • capillary electrophoresis
  • gas chromatography
  • healthcare
  • health information
  • simultaneous determination