IADR Abstract Archives

Human Salivary Proteome Analysis

Human saliva contains a large array of proteins such as enzymes, growth factors, cytokines, immunoglobulins and mucins. Global analysis of the saliva proteins (salivary proteomics) is important for understanding of oral health and disease pathogenesis. Signature proteins in human saliva can also be utilized as diagnostic markers for monitoring human diseases including oral and systematic diseases.

Objective: The study aims to decipher the proteins in human whole saliva by using new proteomics technologies.

Methods: HPLC separation of whole saliva proteins was performed on a C4 reverse-phase column. 35 fractions were collected, and the proteins in each fraction were digested by trypsin for LC-QqTOF MS analysis. Zoom IEF(Invitrogen) and ultracentrifugation were also used to pre-fractionate whole saliva proteins and fractions within different pI or MW ranges were collected for subsequent LC-QqTOF MS analysis. Database search was performed using Mascot database search engine.

A glycoprotein pull-down methodology based on hydrazine chemistry described by Aebersold's group was also demonstrated to identify glycoproteins in human saliva.

Results: In total, 601 distinct proteins were identified from human whole saliva using C4-HPLC/LC-MS/MS, Zoom IEF/LC-MS/MS as well as ultracentrifugation/LC-MS/MS. In addition, 51 glycoproteins were revealed from whole saliva, including 21 novel saliva glycoproteins not found previously.

Conclusion: Both C4-HPLC and Zoom IEF are useful for pre-factionation of human saliva proteome prior to “shotgun” analysis by LC-MS/MS. In total, we have identified 601 distinct saliva proteins, including 51 saliva glycoproteins. Such a global analysis of saliva proteins has provided a “dictionary” for further study of function of saliva proteins as well as discovery of saliva protein biomarkers for human diseases.

*Supported by PHS Grants U01-DE016275 (to D.T.W).


AADR/CADR Annual Meeting
2006 AADR/CADR Annual Meeting (Orlando, Florida)
Orlando, Florida
2006
69
Salivary Research
  • Hu, Shen  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Wong, David T.  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Boontheung, Pinmanee  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Ramachandran, Prasanna  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Xie, Yongming  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Yang, Yanan  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Ieong, Sonya  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Wolinsky, Larry  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Loo, Rachel  ( University of California - Los Angeles, Los Angeles, CA, USA )
  • Loo, Joseph  ( University of California - Los Angeles, Los Angeles, CA, USA )
    • Oral Session
    Salivary Proteomics
    03/08/2006