title: Deleted in colorectal carcinoma (DCC) binds heparin via its fifth fibronectin type III domain.
abstract: DCC (deleted in colorectal carcinoma) is a broadly expressed cell-surface receptor. Netrin-1 was recently identified as a DCC ligand in brain, but the possibility of other DCC ligands was suggested by the finding that an anti-DCC antibody (clone AF5) neutralized netrin-1-dependent commissural axon outgrowth without blocking DCC/netrin-1 interactions. Here we have searched for alternative cell-surface DCC ligands. A DCC-Ig fusion protein bound to neural and epithelial derived cell lines, indicating that these lines express ligand(s) for DCC. The cell-surface binding activity was mediated by the loop between beta-strands F and G of the fifth fibronectin type III repeat FNIII-D5. The loop included the sequence KNRR, which resembles heparin-binding motifs in other proteins. Heparinase and heparitinase treatment of cells reduced binding of DCC-Ig, suggesting that heparan sulfate proteoglycans are cell-surface DCC ligand(s). This was further supported by heparin blocking experiments and by binding of DCC-Ig to immobilized heparan sulfate. The interaction between DCC-Ig and heparan sulfate/heparin, both on the surface of cells and immobilized on plastic, was blocked by the same anti-DCC antibody that blocks netrin-1-dependent commissural axon outgrowth. Taken together, these findings suggest that the DCC-Ig/heparin interaction may contribute to the biological activity of DCC.
authors.affiliation: Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, USA.
authors.forename: K L
authors.initials: KL
authors.lastname: Bennett
authors.forename: J
authors.initials: J
authors.lastname: Bradshaw
authors.forename: T
authors.initials: T
authors.lastname: Youngman
authors.forename: J
authors.initials: J
authors.lastname: Rodgers
authors.forename: B
authors.initials: B
authors.lastname: Greenfield
authors.forename: A
authors.initials: A
authors.lastname: Aruffo
authors.forename: P S
authors.initials: PS
authors.lastname: Linsley
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D015179:Colorectal Neoplasms; D000075389:DCC Receptor; D005353:Fibronectins / Q000737:chemistry*; D016147:Genes, Tumor Suppressor; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D025521:Tumor Suppressor Proteins
chemical_list: D015815:Cell Adhesion Molecules; D000075389:DCC Receptor; C501909:DCC protein, human; D005353:Fibronectins; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D006493:Heparin