Evidence for a cryptic lectin site in the cell-binding domain of plasma fibronectin.
Various proteolytic fragments from the central region of the fibronectin subunit chains containing the main cell-affinity site were applied in cell binding studies using peritoneal macrophages of guinea pigs. A 125I-labelled 23-kDa peptide was relatively well bound by the cells. Attachment to cells was partially inhibited by wheat germ lectin, suggesting a lectin-like site in the cell-binding domain which recognizes oligosaccharide groups with terminal N-acetylglucosamine or N-acetylneuraminic acid. Binding was inhibited by N-acetylneuraminic acid with half-maximal effect at 2 X 10(-3) M. Other inhibitors were a sialic acid rich ganglioside preparation and fetuin, a sialic acid-containing glycoprotein. In contrast to the 23-kDa peptide a 125I-labelled 125-kDa fragment was only weakly bound, although it included the sequence of the 23-kDa peptide on its C-terminus. The residual binding was weakly inhibited by low concentrations of wheat germ lectin and was remarkably improved by higher concentrations. The behavior of the peptide was explained by the presence of a sialic acid-containing oligosaccharide side chain localized outside of the 23-kDa region and interacting with the lectin-like site in the cell-binding sequence. In accord with this suggestion a 95-kDa fragment representing the oligosaccharide-containing part of the 125-kDa peptide was capable of inhibiting at least partially the cell attachment of the 23-kDa piece. The results indicate a lectin-like affinity site in the cell-binding region of fibronectin which is accessible in the 23-kDa peptide, but is masked in the 125-kDa fragment and in fibronectin by a sialic acid-containing oligosaccharide moiety.
D002402:Cathepsin D; D002403:Cathepsins / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D007457:Iodine Radioisotopes; D037102:Lectins / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000097:blood*; D017469:Receptors, Fibronectin; D011975:Receptors, Mitogen / Q000378:metabolism*; D014909:Wheat Germ Agglutinins
D005353:Fibronectins; D007457:Iodine Radioisotopes; D037102:Lectins; D011956:Receptors, Cell Surface; D017469:Receptors, Fibronectin; D011975:Receptors, Mitogen; D014909:Wheat Germ Agglutinins; D002403:Cathepsins; D002402:Cathepsin D