dc.contributor.authorHao, Piliang
dc.contributor.authorGuo, Tiannan
dc.contributor.authorSze, Siu Kwan
dc.date.accessioned2011-05-25T03:08:38Z
dc.date.available2011-05-25T03:08:38Z
dc.date.copyright2011en_US
dc.date.issued2011
dc.identifier.citationHao, P., Guo, T., & Sze, S. K. (2011). Simultaneous Analysis of Proteome, Phospho- and Glycoproteome of Rat Kidney Tissue with Electrostatic Repulsion Hydrophilic Interaction Chromatography. PLoS ONE, 6(2), e16884.en_US
dc.identifier.urihttp://hdl.handle.net/10220/6797
dc.description.abstractProtein post-translational modifications (PTMs) are regulated separately from protein expression levels. Thus, simultaneous characterization of the proteome and its PTMs is pivotal to an understanding of protein regulation, function and activity. However, concurrent analysis of the proteome and its PTMs by mass spectrometry is a challenging task because the peptides bearing PTMs are present in sub-stoichiometric amounts and their ionization is often suppressed by unmodified peptides of high abundance. We describe here a method for concurrent analysis of phosphopeptides, glycopeptides and unmodified peptides in a tryptic digest of rat kidney tissue with a sequence of ERLIC and RP-LC-MS/MS in a single experimental run, thereby avoiding inter-experimental variation. Optimization of loading solvents and elution gradients permitted ERLIC to be performed with totally volatile solvents. Two SCX and four ERLIC gradients were compared in details, and one ERLIC gradient was found to perform the best, which identified 2929 proteins, 583 phosphorylation sites in 338 phosphoproteins and 722 N-glycosylation sites in 387 glycoproteins from rat kidney tissue. Two hundred low-abundance proteins with important functions were identified only from the glyco- or phospho-subproteomes, reflecting the importance of the enrichment and separation of modified peptides by ERLIC. In addition, this strategy enables identification of unmodified and corresponding modified peptides (partial phosphorylation and N-glycosylation) from the same protein. Interestingly, partially modified proteins tend to occur on proteins involved in transport. Moreover, some membrane or extracellular proteins, such as versican core protein and fibronectin, were found to have both phosphorylation and Nglycosylation, which may permit an assessment of the potential for cross talk between these two vital PTMs and their roles in regulation.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesPLoS ONEen_US
dc.rights© 2011 Public Library of Science. This paper was published in PLoS ONE and is made available as an electronic reprint (preprint) with permission of Public Library of Science. The paper can be found at: [DOI: http://dx.doi.org/10.1371/journal.pone.0016884]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.en_US
dc.subjectDRNTU::Science::Biological sciences::Molecular biology
dc.titleSimultaneous analysis of proteome, phospho- and glycoproteome of rat kidney tissue with electrostatic repulsion hydrophilic interaction chromatographyen_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.identifier.doihttp://dx.doi.org/10.1371/journal.pone.0016884
dc.description.versionPublished versionen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record