Saliva is a complex mixture of proteins, lipids and carbohydrates in a physiological buffer containing millimolar concentrations of inorganic phosphate, divalent cations (calcium) and bicarbonate, which supports intermolecular complex formation, tooth mineralization, dental plaque formation, and passive immunity to pathogens of the oral cavity. We propose a four-year proteomics-based research program with the goal to identify and catalogue the components of salivary secretions and to annotate the catalogue with respect to the biochemical properties of the catalogue's inventory. To accomplish high coverage of the proteome, we will implement two parallel approaches to purify and identify individual salivary proteins. First, conventional protein fractionation will be coupled with either multi-dimensional liquid chromatography-tandem electrospray mass spectrometry (LC-MS/MS) or with 2-dimensional geI-MALDI-TOF mass spectrometry. Because the abundant salivary proteins will obscure the purification and identification of medium and low copy salivary proteins, a second purification technology, based on high-throughput immuno-affinity chromatography, will be implemented to complement and support the conventional mass spectrometry-driven proteomics approach. Specifically, we propose to create a comprehensive ? library of anti-saliva antibodies, using bacteriophage display, to build multiple probes for each ? salivary component. These antibody or immune affinity reagents will permit detection, as well as, ? purification of specific salivary proteins, which will, in turn, permit a high-throughput approach to the ? characterization of each protein's post-translational modification state, degree of interaction with ? other salivary components, and affinity for an in vivo tooth model (hydroxyapatite). ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01DE016267-04
Application #
7234130
Study Section
Special Emphasis Panel (ZDE1-PZ (46))
Program Officer
Shum, Lillian
Project Start
2004-09-21
Project End
2009-05-31
Budget Start
2007-06-01
Budget End
2009-05-31
Support Year
4
Fiscal Year
2007
Total Cost
$1,115,545
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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