Automated protein turnover calculations from 15N partial metabolic labeling LC/MS shotgun proteomics data

Author(s)
David Lyon, Maria Angeles Castillejo, Christiana Staudinger, Wolfram Weckwerth, Stefanie Wienkoop, Volker Egelhofer
Abstract

Protein turnover is a well-controlled process in which polypeptides are constantly being degraded and subsequently replaced with newly synthesized copies. Extraction of composite spectral envelopes from complex LC/MS shotgun proteomics data can be a challenging task, due to the inherent complexity of biological samples. With partial metabolic labeling experiments this complexity increases as a result of the emergence of additional isotopic peaks. Automated spectral extraction and subsequent protein turnover calculations enable the analysis of gigabytes of data within minutes, a prerequisite for systems biology high throughput studies. Here we present a fully automated method for protein turnover calculations from shotgun proteomics data. The approach enables the analysis of complex shotgun LC/MS 15N partial metabolic labeling experiments. Spectral envelopes of 1419 peptides can be extracted within an hour. The method quantifies turnover by calculating the Relative Isotope Abundance (RIA), which is defined as the ratio between the intensity sum of all heavy (15N) to the intensity sum of all light (14N) and heavy peaks. To facilitate this process, we have developed a computer program based on our method, which is freely available to download at promex.pph.univie.ac.at/protover.

Organisation(s)
External organisation(s)
Universität Wien
Journal
PLoS ONE
Volume
9
ISSN
1932-6203
DOI
https://doi.org/10.1371/journal.pone.0094692
Publication date
04-2014
Peer reviewed
Yes
Austrian Fields of Science 2012
106005 Bioinformatics, 106037 Proteomics
ASJC Scopus subject areas
Agricultural and Biological Sciences(all), General, Biochemistry, Genetics and Molecular Biology(all)
Portal url
https://ucris.univie.ac.at/portal/en/publications/automated-protein-turnover-calculations-from-15n-partial-metabolic-labeling-lcms-shotgun-proteomics-data(46e1289c-9d1e-483d-b3a7-f906c7b43455).html