Journal article · Book chapter
Exploring extracellular matrix degradomes by TMT-TAILS N-terminomics
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.
Language: | English |
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Publisher: | Humana Press |
Year: | 2019 |
Pages: | 115-126 |
Series: | Methods in Molecular Biology |
ISBN: | 1493990942 , 1493990950 , 9781493990948 and 9781493990955 |
ISSN: | 19406029 and 10643745 |
Types: | Journal article and Book chapter |
DOI: | 10.1007/978-1-4939-9095-5_8 |
ORCIDs: | Madzharova, Elizabeta , Sabino, Fabio and auf dem Keller, Ulrich |