Large-scale phosphoproteomics analysis of whole saliva reveals a distinct phosphorylation pattern

Matthew D. Stone, Xiaobing Chen, Thomas McGowan, Sricharan Bandhakavi, Bin Cheng, Nelson L. Rhodus, Timothy J. Griffin

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

In-depth knowledge of bodily fluid phosphoproteomes, such as whole saliva, is limited. To better understand the whole saliva phosphoproteome, we generated a large-scale catalog of phosphorylated proteins. To circumvent the wide dynamic range of phosphoprotein abundance in whole saliva, we combined dynamic range compression using hexapeptide beads, strong cation exchange HPLC peptide fractionation, and immobilized metal affinity chromatography prior to mass spectrometry. In total, 217 unique phosphopeptides sites were identified representing 85 distinct phosphoproteins at 2.3% global FDR. From these peptides, 129 distinct phosphorylation sites were identified of which 57 were previously known, but only 11 of which had been previously identified in whole saliva. Cellular localization analysis revealed salivary phosphoproteins had a distribution similar to all known salivary proteins, but with less relative representation in "extracellular" and "plasma membrane" categories compared to salivary glycoproteins. Sequence alignment showed that phosphorylation occurred at acidic-directed kinase, proline-directed, and basophilic motifs. This differs from plasma phosphoproteins, which predominantly occur at Golgi casein kinase recognized sequences. Collectively, these results suggest diverse functions for salivary phosphoproteins and multiple kinases involved in their processing and secretion. In all, this study should lay groundwork for future elucidation of the functions of salivary protein phosphorylation.

Original languageEnglish (US)
Pages (from-to)1728-1736
Number of pages9
JournalJournal of Proteome Research
Volume10
Issue number4
DOIs
StatePublished - Apr 1 2011

Keywords

  • dynamic range compression
  • mass spectrometry
  • phosphoproteomics
  • whole saliva

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