Time-resolved analysis of matrix metalloproteinase substrates in complex samples

Pascal Schlage, Fabian E. Egli, Ulrich auf dem Keller*

*Corresponding author for this work

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


Identification of physiological substrates is the key to understanding the pleiotropic functions of matrix metalloproteinases (MMPs) in health and disease. Quantitative mass spectrometry-based proteomics has revolutionized current approaches in protease substrate discovery and helped to unravel many new MMP activities in complex biological systems. Multiplexing further extended the capabilities of these techniques and facilitated more complicated experimental designs that include multiple proteases or monitoring the activity of a single protease at more than one concentration or at multiple time points with a complex test proteome. In this chapter, we provide a protocol for time-resolved iTRAQ-based Terminal Amine Isotopic Labeling of Substrates (TAILS), with the focus on MMP substrate identification and characterization in cell culture supernatants and introduce an automated procedure for the interpretation of time-resolved iTRAQ-TAILS datasets.

Original languageEnglish
Title of host publicationMatrix Metalloproteases
Number of pages14
PublisherHumana Press
Publication date2017
ISBN (Print)978-1-4939-6861-9
ISBN (Electronic)978-1-4939-6863-3
Publication statusPublished - 2017
Externally publishedYes
SeriesMethods in Molecular Biology


  • Protease
  • Proteomics
  • Substrate discovery
  • Time-resolved degradomics

Cite this

Schlage, P., Egli, F. E., & auf dem Keller, U. (2017). Time-resolved analysis of matrix metalloproteinase substrates in complex samples. In Matrix Metalloproteases (Vol. 1579, pp. 185-198). Humana Press. Methods in Molecular Biology, Vol.. 1579 https://doi.org/10.1007/978-1-4939-6863-3_9