Chicken gizzard 5'-nucleotidase functions as a binding protein for the laminin/nidogen complex.
Soluble and reconstituted 5'-nucleotidase were used in the binding assays to the laminin/nidogen complex. They both are shown to interact specifically and in a saturable manner with the laminin/nidogen complex using a solid-phase binding assay. Dissociation constants in the region of 10(-8) M were determined for the association of soluble and membrane-bound 5'-nucleotidase. Scatchard analysis of the binding data indicate a stoichiometry of about 2.7 of the homodimeric soluble 5'-nucleotidase to the laminin/nidogen complex. The association of 5'-nucleotidase with laminin/nidogen occurs in the absence of divalent metal ions and does not require N-linked carbohydrate moieties of both laminin/nidogen and 5'-nucleotidase. 5'-Nucleotidase also associates with isolated laminin although with reduced affinity. No binding to isolated nidogen was observed. Peptides containing the RGD sequence did not influence the binding reaction. Monoclonal and polyclonal antibodies directed against 5'-nucleotidase and laminin specifically perturb the association of the reconstituted enzyme to laminin/nidogen. Sulfated polysaccharides such as heparinsulfate and dermatansulfate modulate the interaction of 5'-nucleotidase and laminin/nidogen in a complex biphasic manner and might also regulate the binding reaction in vivo. Immunohistochemistry shows a close spatial correlation of 5'-nucleotidase and laminin also in the epithelium of the small intestine pointing to an in vivo interaction of both glycoproteins.
Stochaj
U
U
Institut für Zytobiologie und Zytopathologie, Philipps-Universität, Marburg/Deutschland.
Mannherz
H G
HG
D015720:5'-Nucleotidase / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002241:Carbohydrates / Q000502:physiology; D002645:Chickens; D005895:Gizzard, Avian / Q000201:enzymology*; D050356:Lipid Metabolism; D008081:Liposomes / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011510:Proteolipids / Q000378:metabolism; D017474:Receptors, Laminin / Q000378:metabolism*; D015203:Reproducibility of Results; D012995:Solubility
D002241:Carbohydrates; D008081:Liposomes; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011510:Proteolipids; D017474:Receptors, Laminin; C029967:nidogen; C020485:proteoliposomes; D015720:5'-Nucleotidase