These authors contributed equally to this work.
Combined phospho- and glycoproteome enrichment in nephrocalcinosis tissues of phytate-fed rats
Article first published online: 5 NOV 2013
Copyright © 2013 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 27, Issue 24, pages 2767–2776, 30 December 2013
How to Cite
Tran, T., Park, J.-M., Kim, O.-H., Kim, B., Choi, D.-y., Lee, J., Kim, K., Oh, B.-C. and Lee, H. (2013), Combined phospho- and glycoproteome enrichment in nephrocalcinosis tissues of phytate-fed rats. Rapid Commun. Mass Spectrom., 27: 2767–2776. doi: 10.1002/rcm.6742
- Issue published online: 31 OCT 2013
- Article first published online: 5 NOV 2013
- Manuscript Accepted: 18 SEP 2013
- Manuscript Revised: 27 JUL 2013
- Manuscript Received: 11 MAY 2013
Protein post-translational modifications (PTMs) are directly involved in protein function and cellular activities. Among them, glycosylation and phosphorylation are particularly important modifications on proteins located at extracellular and intracellular domains, respectively. However, the combined detection using phospho- and glycoproteomics is limited mainly due to protocol differences.
In this study, we developed a novel method for both phospho- and glycoproteome detection from a single sample batch, in which a titanium dioxide cartridge was used to capture the phosphoproteome, and the flow-through solution was processed for capturing N-linked glycopeptides using hydrazide resin.
By using 1 mg of protein from kidney tissue lysates from normal and diseased rats, we concurrently identified 437 glycosites/358 phosphosites and 468 glycosites/369 phosphosites in normal and disease kidneys, respectively, by liquid chromatography/tandem mass spectrometric analysis.
Compared with individual PTM analyses, the combined PTM analysis clearly provides more broad implications for PTMs related to the pathological status and discovery of biomarker candidates. Furthermore, the combined protocol thoroughly showed its advantages in enrichment efficiency and biological interpretation compared with current methods. Copyright © 2013 John Wiley & Sons, Ltd.