The human salivary proteome is extremely complex, including proteins from salivary glands, serum, and oral microbes. Much has been learned about the host component, but little is known about the microbial component. Here we report a metaproteomic analysis of salivary supernatant pooled from six healthy subjects. For deep interrogation of the salivary proteome, we combined protein dynamic range compression (DRC), multidimensional peptide fractionation, and high-mass accuracy MS/MS with a novel two-step peptide identification method using a database of human proteins plus those translated from oral microbe genomes. Peptides were identified from 124 microbial species as well as uncultured phylotypes such as TM7. Streptococcus, Rothia, Actinomyces, Prevotella, Neisseria, Veilonella, Lactobacillus, Selenomonas, Pseudomonas, Staphylococcus, and Campylobacter were abundant among the 65 genera from 12 phyla represented. Taxonomic diversity in our study was broadly consistent with metagenomic studies of saliva. Proteins mapped to 20 KEGG pathways, with carbohydrate metabolism, amino acid metabolism, energy metabolism, translation, membrane transport, and signal transduction most represented. The communities sampled appear to be actively engaged in glycolysis and protein synthesis. This first deep metaproteomic catalog from human salivary supernatant provides a baseline for future studies of shifts in microbial diversity and protein activities potentially associated with oral disease.
Bibliographical noteFunding Information:
Ken Ashley and Maurice Lirette (British Columbia Ministry of the Environment, Lands and Parks) steered us to Big Lake. John Glew and Dennis Weidler assisted in coring. Drs. Willem Vreeken and Peter Roeder provided tephra analyses, and Kate Laird, Chloë Stuart, Cathy Whitlock, Editors and an anonymous reviewer made helpful comments on earlier versions of this paper. The research was supported by Natural Sciences and Engineering Research Council grants to Brian Cumming, John Smol, and Ian Walker. The paper is dedicated to Julian Szeicz, who lost his life doing fieldwork in northwest Canada.
- Phylogenetic analysis
- Whole saliva