Identification of carboxypeptidase substrates by C-terminal COFRADIC

Sebastian Tanco, Francesc Xavier Aviles, Kris Gevaert, Julia Lorenzo, Petra van Damme

Research output: Chapter in BookChapterResearchpeer-review

2 Citations (Scopus)

Abstract

© Springer Science+Business Media LLC 2017. We here present a detailed procedure for studying protein C-termini and their posttranslational modifications by C-terminal COFRADIC. In fact, this procedure can enrich for both C-terminal and N-terminal peptides through a combination of a strong cation exchange fractionation step at low pH, which removes the majority of nonterminal peptides in whole-proteome digests, while the actual COFRADIC step segregates C-terminal peptides from N-terminal peptides. When used in a differential mode, C-terminal COFRADIC allows for the identification of neo-C-termini generated by the action of proteases, which in turn leads to the identification of protease substrates. More specifically, this technology can be applied to determine the natural substrate repertoire of carboxypeptidases on a proteome-wide scale.
Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages115-133
Number of pages18
Volume1574
DOIs
Publication statusPublished - 1 Jan 2017

Keywords

  • C-terminomics
  • COFRADIC
  • Carboxypeptidases
  • Degradomics
  • Proteases
  • Protein C-termini
  • Substrates

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