This title appears in the Scientific Report :
2017
Please use the identifier:
http://dx.doi.org/10.1007/978-1-4939-6850-3_4 in citations.
Profiling of Protein N-Termini and Their Modifications in Complex Samples
Profiling of Protein N-Termini and Their Modifications in Complex Samples
Protein N termini are a unique window to the functional state of the proteome, revealing translation initiation sites, co-translation truncation and modification, posttranslational maturation, and further proteolytic processing into different proteoforms with distinct functions. As a direct readout...
Saved in:
Personal Name(s): | Demir, Fatih |
---|---|
Niedermaier, Stefan / Kizhakkedathu, Jayachandran N. / Huesgen, Pitter (Corresponding author) | |
Contributing Institute: |
Analytik; ZEA-3 |
Published in: |
Protein Terminal Profiling / Schilling, Oliver (Editor), Chapter 4 |
Imprint: |
New York, NY
Springer New York
2017
|
Physical Description: |
35 - 50 |
ISBN: |
978-1-4939-6849-7 (print) 978-1-4939-6850-3 (electronic) |
DOI: |
10.1007/978-1-4939-6850-3_4 |
PubMed ID: |
28315242 |
Document Type: |
Contribution to a book |
Research Program: |
Plant Science |
Series Title: |
Methods in Molecular Biology
1574 |
Publikationsportal JuSER |
Protein N termini are a unique window to the functional state of the proteome, revealing translation initiation sites, co-translation truncation and modification, posttranslational maturation, and further proteolytic processing into different proteoforms with distinct functions. As a direct readout of proteolytic activity, protein N termini further reveal proteolytic regulation of diverse biological processes and provide a route to determine specific substrates and hence the physiological functions for any protease of interest. Here, we describe our current protocol of the successful Terminal Amine Isotope Labeling of Substrates (TAILS) technique, which enriches protein N-terminal peptides from complex proteome samples by negative selection. Genome-encoded N termini, protease-generated neo-N termini, and endogenously modified N termini are all enriched simultaneously. Subsequent mass spectrometric analysis therefore profiles all protein N termini and their modifications present in a complex sample in a single experiment. We further provide a detailed protocol for the TAILS-compatible proteome preparation from plant material and discuss specific considerations for N terminome data analysis and annotation. |