Enamel extracts induce biomineralization such as osteogenesis and cementogenesis, but the molecular component responsible for this activity remains uncertain. We fractionated enamel extracts from developing pig teeth and isolated the osteoinductive fraction. Proteins from pig enamel scrapings were extracted under alkaline conditions (pH 10.8) and fractionated with the use of a Sephadex G-100 (size exclusion) column. The ability of each fraction to enhance alkaline phosphatase (ALP) activity was assayed in ST2 cells, a mouse bone marrow stromal cell line. The osteoinductive fraction of enamel extracts (OFE) was found in fractions 44 and 45, which induced ST2 cells to express the phenotype of bone-forming osteoblasts, and to form mineralized nodules. Furthermore, the ALP activity of ST2 cells exposed to OFE was reduced by noggin, an antagonist of BMPs, and OFE reacted with BMP-2/4 antibody in dot-blot analysis. These results indicate that OFE contains BMPs that contribute to the induction of biomineralization.