Time-resolved analysis of matrix metalloproteinase substrates in complex samples

Research output: Chapter in Book/Report/Conference proceedingBook chapter – Annual report year: 2017Researchpeer-review

Without internal affiliation

  • Author: Schlage, Pascal

    Swiss Federal Institute of Technology Zurich

  • Author: Egli, Fabian E

    Swiss Federal Institute of Technology Zurich

  • Author: auf dem Keller, Ulrich

    Swiss Federal Institute of Technology Zurich

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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
Volume1579
PublisherHumana Press
Publication date2017
Pages185-198
Chapter9
ISBN (Print)978-1-4939-6861-9
ISBN (Electronic)978-1-4939-6863-3
DOIs
Publication statusPublished - 2017
Externally publishedYes
SeriesMethods in Molecular Biology
Volume1579
ISSN1064-3745
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • iTRAQ-TAILS, Protease, Proteomics, Substrate discovery, Time-resolved degradomics

ID: 140022007