Analysis of the extreme diversity of salivary alpha-amylase isoforms generated by physiological proteolysis using liquid chromatography-tandem mass spectrometry

J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12:911:21-6. doi: 10.1016/j.jchromb.2012.10.023. Epub 2012 Oct 26.

Abstract

Saliva is a crucial biofluid for oral health and is also of increasing importance as a non-invasive source of disease biomarkers. Salivary alpha-amylase is an abundant protein in saliva, and changes in amylase expression have been previously associated with a variety of diseases and conditions. Salivary alpha-amylase is subject to a high diversity of post-translational modifications, including physiological proteolysis in the oral cavity. Here we developed methodology for rapid sample preparation and non-targeted LC-ESI-MS/MS analysis of saliva from healthy subjects and observed an extreme diversity of alpha-amylase proteolytic isoforms. Our results emphasize the importance of consideration of post-translational events such as proteolysis in proteomic studies, biomarker discovery and validation, particularly in saliva.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Biomarkers / analysis
  • Biomarkers / chemistry
  • Biomarkers / metabolism
  • Chromatography, Liquid / methods*
  • Humans
  • Molecular Sequence Data
  • Peptide Fragments / analysis*
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Protein Isoforms
  • Reproducibility of Results
  • Saliva / enzymology*
  • Salivary alpha-Amylases / analysis*
  • Salivary alpha-Amylases / chemistry
  • Salivary alpha-Amylases / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry / methods*
  • Trypsin / metabolism

Substances

  • Biomarkers
  • Peptide Fragments
  • Protein Isoforms
  • Salivary alpha-Amylases
  • Trypsin