title: Endoplasmic reticulum chaperone protein GRP-78 mediates endocytosis of dentin matrix protein 1.
abstract: Dentin matrix protein 1 (DMP1), a phosphorylated protein present in the mineral phase of both vertebrates and invertebrates, is a key regulatory protein during biogenic formation of mineral deposits. Previously we showed that DMP1 is localized in the nuclear compartment of preosteoblasts and preodontoblasts. In the nucleus DMP1 might play an important role in the regulation of genes that control osteoblast or odontoblast differentiation. Here, we show that cellular uptake of DMP1 occurs through endocytosis. Interestingly, this process is initiated by DMP1 binding to the glucose-regulated protein-78 (GRP-78) localized on the plasma membrane of preodontoblast cells. Binding of DMP1 to GRP-78 receptor was determined to be specific and saturable with a binding dissociation constant K(D)=85 nm. We further depict a road map for the endocytosed DMP1 and demonstrate that the internalization is mediated primarily by caveolae and that the vesicles containing DMP1 are routed to the nucleus along microtubules. Immunohistochemical analysis and binding studies performed with biotin-labeled DMP1 confirm spatial co-localization of DMP1 and GRP-78 in the preodontoblasts of a developing mouse molar. Co-localization of DMP1 with GRP-78 was also observed in T4-4 preodontoblast cells, dental pulp stem cells, and primary preodontoblasts. By small interfering RNA techniques, we demonstrate that the receptor for DMP1 is GRP-78. Therefore, binding of DMP1 with GRP-78 receptor might be an important mechanism by which DMP1 is internalized and transported to the nucleus during bone and tooth development.
authors.affiliation: Department of Oral Biology, University of Illinois, Chicago, Illinois 60612, USA.
authors.forename: Sriram
authors.initials: S
authors.lastname: Ravindran
authors.forename: Karthikeyan
authors.initials: K
authors.lastname: Narayanan
authors.forename: Asha Sarah
authors.initials: AS
authors.lastname: Eapen
authors.forename: Jianjun
authors.initials: J
authors.lastname: Hao
authors.forename: Amsaveni
authors.initials: A
authors.lastname: Ramachandran
authors.forename: Sylvie
authors.initials: S
authors.lastname: Blond
authors.forename: Anne
authors.initials: A
authors.lastname: George