Bone and dentin formation are interesting examples of matrix-mediated mineralization. However, factors and mechanisms regulating this process are poorly understood. Dentin matrix protein 1 (DMP1) is an acidic extracellular matrix protein found in dentin and bone, and based on its amino acid composition it could be postulated to play an important role in mineralization. Our present study examines the ability of recombinant DMP1 to initiate apatite formation in vitro. A 45 Ca-binding assay demonstrated that recombinant DMP1 (rDMP1) possesses calcium-binding ability under physiological conditions. The in vitro nucleation experiments when conducted with rDMP1-coated glass plates demonstrated hydroxyapatite nucleation, while amorphous mineral was deposited on blank or BSA-coated surface. This mineral deposition was found to be 10-fold higher on rDMP1-coated glass surface when compared with the control glass plates. These findings suggest that DMP1 could be considered as a nucleator for apatite deposition in vitro.