Ameloblastin Regulates Osteogenic Differentiation by Inhibiting Src Kinase via Cross Talk between Integrin β1 and CD63

Abstract

Ameloblastin, the most abundant nonamelogenin enamel matrix protein, plays a role in ameloblast differentiation. Here, we found that ameloblastin was expressed in osteosarcoma cells; to explore the potential functions of ameloblastin in osteoblasts, we investigated whether this protein is involved in osteogenic differentiation and bone formation on the premise that CD63, a member of the transmembrane-4 glycoprotein superfamily, interacts with integrins in the presence of ameloblastin. Ameloblastin bound to CD63 and promoted CD63 binding to integrin β1. The interaction between CD63 and integrin β1 induced Src kinase inactivation via the binding of CD63 to Src. The reduction of Src activity and osteogenic differentiation mediated by ameloblastin were abrogated by treatment with anti-CD63 antibody and overexpression of constitutively active Src, respectively. Therefore, our results suggest that ameloblastin is expressed in osteoblasts and functions as a promoting factor for osteogenic differentiation via a novel pathway through the interaction between CD63 and integrin β1.

ACKNOWLEDGMENTS

We thank Nanci Antonio (Université de Montréal) for helpful discussions. We also thank Saki Iizuka and Atsuhiro Nagasaki (Hiroshima University) for technical assistance. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan (to T.T., Y.K., M.M., and I.O.) and a Research Fellowship for Young Scientists and Excellent Young Researchers Overseas Visit Program from the Japan Society for the Promotion of Science (to S.I.).

We declare that we have no conflicts of interest.