Isolation and use fibronectin receptor

Details Isolation and use fibronectin receptor Isolation and use fibronectin receptor

1998-05-20

2001-06-05

Saunders, David (Department: 1644)

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

Blood proteins or globulins, e.g., proteoglycans, platelet...

C530S389100

Reexamination Certificate

active

06242577

ABSTRACT:

FIELD OF THE INVENTION
This invention relates generally to the field of biochemistry and more particularly to a cell surface glycoprotein having the apparent molecular weight of 140,000 daltons with the properties expected of a fibronectin receptor, and the use of the receptor to prepare liposomes with predetermined adhesion properties.
BACKGROUND OF THE INVENTION
Cell-substrate adhesion is generally considered to be a multistep process involving recognition of extracellular matrix components by cell surface receptors, followed by cytoskeletal rearrangements that lead to cell spreading (Grinnell, 1978; Hynes, 1981). Several extracellular matrix glycoproteins, such as fibronectin (Ruoslahti, et al., 1981b), laminin (Timpl, et al., 1979), vitronectin (Hayman, et al., 1983), and collagens have been shown to promote attachment of various cell types to tissue culture substrates (Kleinman, et al., 1981). The cell membrane receptors that recognize these matrix proteins, however, remain essentially unknown, although putative receptors for laminin (Lesot, et al., 1983; Malinoff and Wicha, 1983) and collagens (Chiang and Kang, 1982; Mollenhauer and von der Mark, 1983) are currently being investigated.
A number of candidates for the role of a fibronectin receptor have been proposed. By photoaffinity labeling, it was shown that a 49 kd glycoprotein comes into close contact with substrate-bound fibronectin (Aplin, et al., 1981). Further support for the notion that the receptor is a protein comes from studies showing that treatment of cells with certain proteases abolishes the ability of cells to attach to fibronectin (Tarone, et al., 1982). Treatment with trypsin, however, at least in the presence of Ca
++
, leaves the receptor activity intact (Oppenheimer-Marks and Grinnell, 1984). Based upon the calcium-dependent stability to trypsin, Oppenheimer-Marks and Grinnell (1984) have proposed a 48 kd wheat germ agglutinin-binding glycoprotein as a potential fibronectin receptor.
It has also been suggested that heparan sulfate proteoglycans might be involved in cell attachment to fibronectin (Culp, et al., 1979; Laterra, et al., 1983). Indeed, photocross-linking experiments performed by Perkins, et al. (1979) showed that proteoglycans are associated with fibronectin at the cell surface.
A different type of cell surface component has been implicated in fibronectin-cell interactions by studies showing an inhibitory effect of di- and trisialogangliosides on the attachment of cells to fibronectin (Kleinman, et al., 1979). The inhibitory activity was found to reside in the carbohydrate moiety of the glycolipid. Antibodies that interfere with cell attachment have been described by a number of investigators, and the corresponding antigens have been found to be proteins with molecular weights ranging from 60 to 160 kd (Hsieh and Sueoka, 1980; Knudsen, et al., 1981; Neff, et al., 1982; Greve and Gottlieb, 1982; Oesch and Birchmeier, 1982), or specific gangliosides (Dippold, et al., 1984).
A large number of binding affinities are known to be present in the fibronectin molecule, such as for collagen (Engvall and Ruoslahti, 1977), fibrinogen and fibrin (Ruoslahti and Vaheri, 1975), proteoglycans (Stathakis and Mosesson, 1977), cell surfaces (Klebe, 1974; Pearlstein, 1976), and actin (Keski-Oja, et al., 1980), and there have been some studies of the interaction of cell surfaces with the cell attachment site. (Pierschbacher, et al., 1981). A large fibronectin fragment, that promotes cell attachment but lacks the other binding activities is also known, (Pierschbacher, et al., 1982, 1983; Pierschbacher and Ruoslahti, 1984a).
It has now been discovered that a 140 kd protein from detergent extracts of cells, when incorporated into liposomes, promotes their binding specifically to fibronectin-coated substrates via the Arg-Gly-Asp sequence in the fibronectin molecule.
REFERENCES
The content of the following references is incorporated into the foregoing specification, as fully as though set forth therein as a background for those skilled in the art.
Aplin, J. D., Hughes, R. C., Jaffe, C. L., and Sharon, N. (1981) Reversible cross-linking of cellular components of adherent fibroblasts to fibronectin and lectin-coated substrata. Exp. Cell Res. 134, 488-494.
Billiau, A., Edy, V. G., Heremans, H., Van Damme, J., Desmyter, J., Georgiades, J. A., and DeSomer, P. (1977). Human interferon: mass production in a newly established cell line, MG-63. Antimicrob. Agents Chemother. 12, 11-15.
Cheresh, D. A., Harper, J. R., Schulz, G., and Reisfeld, R. A. (1984). Localization of the gangliosides GD
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and GD
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Culp. L. A., Murray, B. A., and Rollins, B. J. (1979). Fibronectin and proteoglycans as determinants of cell-substratum adhesion. J. Supramol. Struct. 11, 401-427.
Dippold, W. G., Knuth, A., and Meyer zum Büschenfelde, K. (1984). Inhibition of human melanoma cell growth in vitro by monoclonal anti-GD3-ganglioside antibody. Cancer Res. 44, 806-810.
Engvall, E., and Ruoslahti, E. (1977). Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int. J. Cancer 20, 1-5.
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Greve, J. M., and Gottieb, D. I. (1982). Monoclonal antibodies which alter the morphology of cultured chick myogenic cells. J. Cell. Biochem. 18, 221-229.
Grinnell, F. (1978). Cellular adhesiveness and extracellular substrata. Int. Rev. Cyto. 53, 65-144.
Hayman, E. G., Engvall, E., and Ruoslahti, E. (1981). Concomitant loss of cell surface fibronectin and laminin from transformed rat kidney cells. J. Cell Biol. 88, 352-357.
Hayman, E. G., Pierschbacher, M. D., Ohgren, Y., and Ruoslahti, E. (1983). Serum spreading factor (vitronectin) is present at the cell surface and in tissues. Proc. Nat. Acad. Sci. USA 80, 4003-4007.
Hoffman, S., Sorkin, B. C., White, P. C., Brackenbury, R., Mailhammer, R., Rutishauser, U., Cunningham, B. A., and Edelman, G. M. (1982). Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J. Biol. Chem. 257, 7720-7729.
Hsieh, P., and Sueoka, N. (1980). Antisera inhibiting mammalian cell spreading and possible cell surface antigens involved. J. Cell Biol. 86, 866-873.
Hynes, R. O. (1981). Relationships between fibronectin and the cytoskeleton. In Cell Surface Reviews, Vol. 7, G. Poste and G. L. Nicolson, eds (Amsterdam: Elsevier/North Holland), pp. 97-136.
Keski-Oja, J., Sen. A., and Todaro, G. S. (1980). Direct association of fibronectin and actin molecules in vitro. J. Cell Biol. 85, 527.
Klebe, R. J. (1974). Isolation of a collagen-dependent cell attachment factor. Nature 250, 248-251.
Kleinman, H. K., Martin., G. R., and Fishman, P. H. (1979). Ganglioside inhibition of fibronectin mediated cell adhesion to collagen. Proc. Nat. Acad. Sci. USA 76, 3367-3371.
Kleinman, H. K., Klebe, R. J., and Martin, G. R. (1981). Role of collagenous matrices in the adhesion and growth of cells. J. Cell Biol. 88, 473-485.
Knudsen, K. A., Rao, P. E., Damsky, C. H., and Buck, C. A. (1981). Membrane glycoproteins involved in cell-substratum adhesion. Proc. Nat. Acad. Sci. USA 78, 6071-6075.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.
Laterra, J., Siebert, J. E., and Culp, L. A. (1983). Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. J. Cell Biol. 96, 112-123.
Lebien, T. W., Boue, D. R., Bradley, J. G., and Kersey, J. H. (1982). Antibody affinity may influence antigenic modulation of the common acute lymphoblastic leukemia antigen in vitro. J. Immunol. 129, 2287-2292

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