Chapter title |
Time-Resolved Analysis of Matrix Metalloproteinase Substrates in Complex Samples
|
---|---|
Chapter number | 9 |
Book title |
Matrix Metalloproteases
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6863-3_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6861-9, 978-1-4939-6863-3
|
Authors |
Pascal Schlage, Fabian E. Egli, Ulrich auf dem Keller |
Editors |
Charles A. Galea |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 20 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 4 | 20% |
Researcher | 2 | 10% |
Student > Master | 2 | 10% |
Professor | 2 | 10% |
Unspecified | 1 | 5% |
Other | 2 | 10% |
Unknown | 7 | 35% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 7 | 35% |
Unspecified | 1 | 5% |
Chemical Engineering | 1 | 5% |
Mathematics | 1 | 5% |
Agricultural and Biological Sciences | 1 | 5% |
Other | 3 | 15% |
Unknown | 6 | 30% |