Potential biomarkers of human salivary function: a modified proteomic approach.
OBJECTIVE
In previous studies, we defined groups of subjects with opposite salivary function. Group membership was associated with clinically relevant outcomes. High aggregation-adherence (HAA) groups showed lower levels of caries, supragingival plaque, total streptococci, and Tannerella forsythensis than low high aggregation-adherence (LAA) groups. In this study, we used a proteomic approach to search for biomarkers which could be useful as risk indicators for those outcomes.


METHODS
Clarified resting whole saliva from each of 41 HAA and LAA subjects was separated by preparative isoelectric focusing. Fractions showing the most distinctive protein profiles were pooled into four sets (pI 3-3.5, pI 4-4.7, pI 5.7-7.7, pI 10-11.5). Each pool then was compared by SDS-PAGE. Image analysis software was used to quantify matched bands. Partial least squares analysis (PLS) was used to determine which of the 65 bands from all four pools were the best predictors of group membership, caries, total plaque, total streptococci, and T. forsythensis counts. Those bands were identified by mass spectroscopy (MS-MS).


RESULTS
Two bands consistently were strong predictors in separate PLS analyses of each outcome variable. In follow-up univariate analyses, those bands showed the strongest significant differences between the HAA and LAA groups. They also showed significant inverse correlations with caries and all the microbiological variables. MSMS identified those bands as statherin, and a truncated cystatin S missing the first eight N-terminal amino acids.


CONCLUSIONS
Levels of statherin and truncated cystatin S may be potential risk indicators for the development of caries and other oral diseases.
Rudney
J D
JD
Department of Diagnostic and Biological Sciences, School of Dentistry, 17-252 Moos Tower, 515 Delaware St. SE, University of Minnesota, Minneapolis, MN 55455, USA. jrudney@tc.umn.edu
Staikov
R K
RK
Johnson
J D
JD
D001422:Bacterial Adhesion / Q000502:physiology; D018441:Biofilms; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D015169:Colony Count, Microbial; D003731:Dental Caries / Q000382:microbiology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D040901:Proteomics / Q000379:methods; D012307:Risk Factors; D012463:Saliva / Q000737:chemistry* / Q000382:microbiology / Q000502:physiology; D055333:Salivary Cystatins / Q000032:analysis*; D012471:Salivary Proteins and Peptides / Q000032:analysis*
D015415:Biomarkers; C018149:STATH protein, human; D055333:Salivary Cystatins; D012471:Salivary Proteins and Peptides