Self-assembly of a recombinant amelogenin protein generates supramolecular structures.
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Amelogenin proteins are the principal constituents of the extracellular organic matrix associated with the nucleation and growth of the carbonated calcium hydroxyapatite (HAP)-containing mineral phase of dental enamel. Amelogenins are believed to function in controlling the sizes and organization of the developing enamel crystals. Previous studies have shown that enamel proteins exhibit unusual reversible aggregation properties. The present studies were designed to test the hypothesis that self-assembly of recombinant amelogenin generates supramolecular structures that are indistinguishable from the electron-dense particles associated with HAP crystal growth in vivo. A recombinant amelogenin analog of the murine 180-residue protein was analyzed by high-resolution size exclusion chromatography, atomic force (AFM), and transmission electron (TEM) microscopy. It was found that the amelogenin formed supramolecular aggregates which were in a concentration-dependent equilibrium with protein monomers. Imaging of the amelogenin by both AFM and TEM techniques revealed spherical aggregate structures of about 18 nm diameter which were seen to be similar to electron-dense enamel structures observed in vivo. We interpret these results to suggest that, in vivo, the amelogenin protein self-assembles through functional motifs of the protein primary structure, generating specific supramolecular aggregates which we hypothesize function to control the ultrastructural organization of the developing enamel crystallites.