Two classes of dentin phosphophoryns, from a wide range of species, contain immunologically cross-reactive epitope regions.

An immunological species comparison, using a monospecific rabbit polyclonal antibody directed against rat incisor alpha-phosphophoryn, has been undertaken to assess the similarity in epitope regions among various dentin phosphophoryns (PP) that were prepared from human, monkey, bovine, ovine, and echinoderm teeth. Dentin extracellular matrix proteins were extracted with a standard method using 0.5 M EDTA in the presence of enzyme inhibitors. Final phosphophoryn purification was performed on DEAE ion exchange HPLC. Cross-reactivity of the polyclonal antibody was examined by enzyme-linked immunosorbant assay (ELISA) and dot-blot. The results of this investigation demonstrate a cross-reactivity of the rat-alpha-phosphophoryn antibody (anti-RIPP) with at least one phosphophoryn component in each dentin studied, indicating the existence of similar antigenic determinants among these proteins. It would seem that these epitope regions have been strongly conserved since the epitope region is also present in the phosphoprotein of echinoderm teeth. No cross-reactivity was found with phosvitin (a phosphoserine-rich phosphoprotein), rat serum albumin, bovine serum albumin, or collagen type IV. However, a new and distinct second cross-reactive phosphophoryn, not calcium ion-precipitable, was found in the EDTA insoluble fraction from the teeth. These results indicate that dentin phosphophoryns are specific phenotypic markers for odontoblast expression. Because of the species cross-reactivity, the polyclonal anti-RIPP antibody may be a useful probe in studying the distribution of phosphophoryns in other species, such as human teeth.