Osteopontin (OPN) and osteonectin/SPARC (ON/SPARC) are prominent matricellular components of the extracellular matrix of mineralized tissues of bones and teeth in which they can regulate the formation and growth of hydroxyapatite crystals and influence a variety of cell activities. OPN regulates cell responses through several integrin receptors and is also a ligand for the CD44 receptor, through which it acts as a chemoattractant. Although a cell-surface receptor for SPARC has not been identified it can block cell-cell and cell-matrix interactions and inhibit cell migration and chemotaxis. OPN and SPARC also appear to function inside cells. Thus, OPN appears to exist in association with the CD44 receptor inside migratory cells, while intracellular SPARC is associated with axonemal tubulin in ciliated epithelial cells. Analyses of fibroblasts and peritoneal macrophages from OPN-null and CD44-null cells show impaired functionality involving migration and cell fusion required for osteoclast formation, while disruption of SPARC expression leads to developmental defects in Xenopus. To gain further insights into the intracellular functions of OPN and SPARC, we have used the yeast two-hybrid system to identify potential interacting molecules. Using full-length SPARC as bait the carboxy-terminal domain, which contains two EF-hand, high-affinity binding sites, was found to have transcriptional activity, while several novel proteins that interact with the amino-terminal domains of SPARC and full-length OPN have been identified. The identification of OPN and SPARC inside specialized cells introduces a novel concept in cellular regulation by matricellular proteins.