title: Dentin matrix protein-1 isoforms promote differential cell attachment and migration.
abstract: Dentin matrix protein-1 (DMP1), bone sialoprotein (BSP), and osteopontin (OPN) are three SIBLINGs (small integrin-binding ligand, N-linked glycoproteins) co-expressed/secreted by skeletal and active ductal epithelial cells. Although etiological mechanisms remain unclear, DMP1 is the only one of these three genes currently known to have mutations resulting in human disease, and yet it remains the least studied. All three contain the highly conserved integrin-binding tripeptide, RGD, and experiments comparing the cell attachment and haptotactic migration-enhancing properties of DMP1 to BSP and OPN were performed using human skeletal (MG63 and primary dental pulp cells) and salivary gland (HSG) cells. Mutation of any SIBLING's RGD destroyed all attachment and migration activity. Using its alphaVbeta5 integrin, HSG cells attached to BSP but not to DMP1 or OPN. However, HSG cells could not migrate onto BSP in a modified Boyden chamber assay. Expression of alphaVbeta3 integrin enhanced HSG attachment to DMP1 and OPN and promoted haptotactic migration onto all three proteins. Interchanging the first four coding exons or the conserved amino acids adjacent to the RGD of DMP1 with corresponding sequences of BSP did not enhance the ability of DMP1 to bind alphaVbeta5. For alphaVbeta3-expressing cells, intact DMP1, its BMP1-cleaved C-terminal fragment, and exon six lacking all post-translational modifications worked equally well but the proteoglycan isoform of DMP1 had greatly reduced ability for cell attachment and migration. The sequence specificity of the proposed BMP1-cleavage site of DMP1 was verified by mutation analysis. Direct comparison of the three proteins showed that cells discriminate among these SIBLINGs and among DMP1 isoforms.
authors.affiliation: Craniofacial and Skeletal Diseases Branch, NIDCR, National Institutes of Health, Bethesda, Maryland 20892-4320, USA.
authors.forename: Zofia
authors.initials: Z
authors.lastname: von Marschall
authors.forename: Larry W
authors.initials: LW
authors.lastname: Fisher
mesh_terms: D020816:Amino Acid Motifs; D000596:Amino Acids / Q000737:chemistry; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D005091:Exons; D016326:Extracellular Matrix Proteins / Q000737:chemistry*; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000737:chemistry; D009154:Mutation; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry*; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary
chemical_list: D000596:Amino Acids; C513309:DMP1 protein, human; D016326:Extracellular Matrix Proteins; D039302:Integrin alphaVbeta3; D009842:Oligopeptides; D010455:Peptides; D010750:Phosphoproteins; D020033:Protein Isoforms; C047981:arginyl-glycyl-aspartic acid