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

Therese Dau, Kapil Gupta, Imre Berger, Juri Rappsilber*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

22 Citations (Scopus)
220 Downloads (Pure)


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.

Original languageEnglish
Pages (from-to)4472-4478
Number of pages7
JournalAnalytical Chemistry
Issue number7
Early online date28 Feb 2019
Publication statusPublished - 2 Apr 2019

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute


  • Synthetic biology


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