| 1557 Proteomic analysis of mice salivary proteins adsorbed on hydroxyapatite (HAP) | ||
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K. OSHIMA, Tsurumi University, Ichikawa, Chiba, Japan, T. OHSHIMA, Tsurumi University, Yokohama, Japan, and N. MAEDA, Tsurumi University, School of Dental Medicine, Yokohama City, Japan Objectives: We have already reported that the caries susceptibility of BALB/cA mice was higher than that of C3H/HeN. Salivary proteins might have an important role to develop dental caries. In order to clarify the difference of caries susceptibility in 2 different inbred mouse strains, we carried out the proteomic analysis of salivary proteins obtained from BALB/cA and C3H/HeN mice and compared between them. Methods: Collection of saliva samples: The whole saliva was colleted from BALB/cA and C3H/HeN by stimulating with pilocarpin sulfate and isoproterenol hydrochloride and pooled. HAP-HPLC: The whole saliva sample was eluted and fractionated with HAP-HPLC linear- gradient from 10mM to 0.5M of potassium phosphate buffer. Each fraction was desalted and separated by SDS-PAGE and visualized by silver staining. Protein electrophoresis and peptide mass-fingerprinting: An aliquot of saliva was incubated with HAP beads of 0.5mm diameter at 37C for 30 minutes on the rotator, and removed the free proteins by washing with saline. The adsorbed proteins were eluted with 0.5M potassium phosphate buffer (pH6.8), desalted, and separated by 12.5%SDS-PAGE or two-dimensional SDS-PAGE and CBB stain was carried out. The different spots were picked up, cleaved by Trypsin, and analysed by Mass-Spectrometer. Results: The most major salivary proteins of two strains adsorbed to HAP were different in the mobility of 12.5% SDS-PAGE around 45KDa. The different spots in 2D-PAGE corresponding to 45KDa bands of SDS-PAGE were revealed as Carbonic-anhydrase family by peptide mass fingerprinting and BLAST search. The result of HAP-HPLC analysis revealed the fractions around 40 and 15KDa were unique for BALB/cA, and 50 and 30KDa were unique for C3H/HeN. Conclusions: These results suggested that the differences of plural salivary proteins adsorbed on HAP in BALB/cA and C3H/HeN mice might associate with the caries susceptibility in these mouse strains. | ||
| Seq #192 - Salivary Proteomics 10:45 AM-12:45 PM, Friday, 11 March 2005 Baltimore Convention Center 322-323 | ||
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