title: Non-RGD domains of osteopontin promote cell adhesion without involving alpha v integrins.
abstract: Osteopontin (OPN) is an integrin-binding secreted protein that contains an Arg-Gly-Asp (RGD) amino acid sequence and binds to various cell types via RGD-mediated interaction with the alpha v beta 3 integrin. We have identified a cell line whose binding to OPN does not require RGD or alpha v interactions. We compared the ability of two murine cell lines, L929 fibroblastic cells and B16-BL6 melanoma cells, to interact with OPN (from human milk, and recombinant human and mouse OPN) as well as recombinant OPN prepared to include either the N-terminal or C-terminal halves but lacking the RGD sequence. Both cell lines adhered to GRGDS peptides coupled to BSA, and these interactions were inhibited by addition of GRGDS (but not GRGES) peptides or a monoclonal antibody specific to the alpha v integrin subunit. Adhesion of L929 cells to OPN was also dependent on the RGD sequence and the alpha v integrin subunit. However, the binding of B16-BL6 cells was not inhibited by either GRGDS peptides or the anti-alpha v antibody. B16-BL6 (but not L929) cells were also able to adhere to and spread on both N-terminal and C-terminal OPN proteins that lack the RGD sequence, and these interactions were not inhibited by either GRGDS peptides or anti-alpha v antibody. Together these results indicate that B16-BL6 cells can adhere to OPN by interactions that are independent of either the RGD sequence or the alpha v integrin subunit, and suggest that some cells can interact with additional, non-RGD binding sites in OPN.
authors.affiliation: Section of Immunopathogenesis, Hokkaido University, Sapporo, Japan.
authors.forename: Y U
authors.initials: YU
authors.lastname: Katagiri
authors.forename: M
authors.initials: M
authors.lastname: Murakami
authors.forename: K
authors.initials: K
authors.lastname: Mori
authors.forename: J
authors.initials: J
authors.lastname: Iizuka
authors.forename: T
authors.initials: T
authors.lastname: Hara
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: W Y
authors.initials: WY
authors.lastname: Jia
authors.forename: A F
authors.initials: AF
authors.lastname: Chambers
authors.forename: T
authors.initials: T
authors.lastname: Uede
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D002460:Cell Line; D002465:Cell Movement; D006801:Humans; D039564:Integrin alphaV; D008545:Melanoma / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D053495:Osteopontin; D011956:Receptors, Cell Surface / Q000378:metabolism; D012795:Sialoglycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D039564:Integrin alphaV; D009842:Oligopeptides; D011956:Receptors, Cell Surface; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D053495:Osteopontin; C047981:arginyl-glycyl-aspartic acid