Proteome analysis of high affinity mouse saliva proteins to hydroxyapatite. Issue 8 (August 2022)

• Record Type: Journal Article
• Authors: Ohshima, Keijiro; Ohshima, Tomoko; Meyer, Karen; Takai, Eisuke; Yoshizawa, Shunsuke; Shiraki, Kentaro; Maeda, Nobuko
• Is Part Of: Heliyon. Volume 8:Issue 8(2022)
• Publication Date: 2022-08
• Subjects: Mouse saliva -- Hydroxyapatite -- Dental caries -- Carbonic anhydrase VI; Research -- Periodicals; Medical sciences -- Periodicals; Natural history -- Periodicals; Social sciences -- Periodicals; Earth sciences -- Periodicals; Physical sciences -- Periodicals; 507.2
• DOI: 10.1016/j.heliyon.2022.e10077
• Languages: English
• ISSNs: 2405-8440

Abstract

Abstract: Caries sensitivity varies between the two strains of inbred mice, BALB/cA has high sensitivity and C3H/HeN has low sensitivity. One potential reason seems to be a difference in pellicle-forming saliva protein composition. Here, we performed a proteomic analysis in order to identify differences of hydroxyapatite (HAP) adsorbed saliva proteins between these two mouse strains. HAP column chromatography revealed twice the quantity of high-affinity saliva proteins in C3H/HeN compared to BALB/cA. One- and two-dimensional electrophoresis showed 2 bands/spots with deviating migration. They were identified as murine carbonic anhydrase VI (CAVI) by peptide mass fingerprinting and confirmed with western blotting using a specific polyclonal antibody. Total RNA from the salivary glands of both mouse strains, PCR amplification of cDNA with a CAVI specific primer, and sequence analysis revealed one different base in codon 96, resulting in one different amino acid. Glyco-chains of CAVI deviate in one N-glycan, confirmed by mass analysis. CAVI activity was estimated from distinct circular dichroism spectra of the molecules and found higher in C3H/HeN mice. In summary, the CAVI composition of BALB/cA and C3H/HeN differs in one amino acid and a glyco-chain modification. Further, saliva from caries resistant C3H/HeN mice displayed higher CAVI activity and also overall hydroxyapatite adsorption, suggesting a relationship with caries susceptibility. Highlights: CAVI was the salivary protein with high affinity for hydroxyapatite in two mice strains with different caries susceptibility. CAVI of the two strains showed differences in molecular weight, amino acids and genes, glyco-chain modification and enzyme activity. Differences in CAVI activity might contribute to caries susceptibility. Abstract : Mouse saliva; Hydroxyapatite; Dental caries; Carbonic anhydrase VI