TOOTH ENAMELINS IDENTIFIED MAINLY AS SERUM-PROTEINS - MAJOR ENAMELIN IS ALBUMIN

The major enamelin protein component present in EDTA or EDTA/guanidine hydrochloride extracts of developing bovine enamel has a molecular mass of about 67 kDa; it has an amino acid composition similar to that of bovine serum albumin and reacts with polyclonal and monoclonal antibodies to albumin. Two‐dimensional separation of the components in the enamelin extract by isoelectric focusing and SDS/PAGE reveal that the major ∼ 67‐kDa component and almost all of the minor Coomassie‐staining protein components of 7ap; 67 kDa, as well as many of the other minor components with different molecular masses, also react with polyclonal and monoclonal antialbumin. The ∼ 67‐kDa band eluted after SDS/PAGE, as well as the major ∼ 67‐kDa spots eluted after two‐dimensional separation, were found to have N‐terminal amino acid sequences identical to that of bovine serum albumin. Albumin accounted for at least 70–80% of the total protein content of the enamelin extract and was essentially the only protein in the ∼ 67‐kDa component. The serum proteins α‐2 HS glycoprotein, γ‐globulin and fetuin, and the proline‐rich salivary protein termed P‐B were also identified in the enamelin extract. The serum proteins and the salivary protein account for > 95% of the proteins in the enamelin extracts. Of the remaining very small amounts of non‐serum or salivary protein isolated from the enamelin extracts, three minor components were isolated which had N‐terminal amino acid sequences which were not similar to any known protein in the protein sequence data base and could therefore conceivably be true ‘enamelins’ synthesized by ameloblasts. One additional protein had the first five N‐terminal amino acids and residue 8 of amelogenin, residues 6 and 7 being different from those of amelogenin. Two other very minor protein components had amino acid compositions distinct from the amelogenins and the serum proteins, but were N‐terminally blocked on attempted sequencing. None of the components in the neutral soluble low‐ionic‐strength extract or in the 4 M guanidine hydrochloride extract, both of which consist principally of amelogenins, immunoreacted with anti‐albumin or with any of the antibodies to other serum proteins and fetuin, despite the fact that the amelogenin extracts also contain non‐amelogenin proteins. On the basis of the data presented, studies employing antibodies to the so‐called enamelin proteins and hypotheses as to their molecular conformation, their roles as evolutionary markers, or their positive role in mineralization should be reconsidered and reviewed. Copyright © 1990, Wiley Blackwell. All rights reserved