Exploring extracellular matrix degradomes by TMT-TAILS N-terminomics

Elizabeta Madzharova, Fabio Sabino, Ulrich auf dem Keller*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

Global characterization of protein N termini provides valuable information on proteome dynamics and diversity in health and disease. Driven by the progress in mass spectrometry-based proteomics, novel approaches for the dedicated investigation of protein N termini and protease substrates have been recently developed. Terminal amine isotopic labeling of substrates (TAILS) is a quantitative proteomics approach suitable for high-throughput and system-wide profiling of protein N termini in complex biological matrices. TAILS employs isotopic labeling of primary amines of intact proteins in combination with an amine-reactive high molecular weight polymer (HPG-ALD) for depletion of internal tryptic peptides and high enrichment of protein N termini by negative selection. Thereby, TAILS allows simultaneous identification of the natural N termini, protease-generated neo-N termini, and endogenously modified (e.g., acetylated) N termini. In this chapter, we provide a protocol for tandem mass tag (TMT)-TAILS analysis and further discuss specific considerations regarding N-terminome data interpretation using Proteome Discoverer™ software.

Original languageEnglish
Title of host publicationCollagen
EditorsIrit Sagi, Nikolaos A. Afratis
Number of pages12
PublisherHumana Press
Publication date2019
Pages115-126
Chapter8
ISBN (Print)978-1-4939-9094-8
ISBN (Electronic)978-1-4939-9095-5
DOIs
Publication statusPublished - 2019
SeriesMethods in Molecular Biology
Volume1944
ISSN1064-3745

Keywords

  • N-terminome
  • Proteolysis
  • Proteome Discoverer™
  • TAILS
  • TMT

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