Lactoferrin-binding sites at the surface of HT29-D4 cells. Comparison with transferrin.
The binding of 125I-lactoferrin to HT29-D4 cells, a clone of HT29 cells, was studied and compared to the binding of 125I-transferrin to the same cells. The binding of the two iron-transport proteins is saturable and reversible suggesting the presence of specific receptors for each protein. Scatchard analysis suggests the existence of binding sites for lactoferrin with the relatively high equilibrium dissociation constant, Kd1 of 408 nM. Additionally, the cell is capable of binding large amounts of lactoferrin with very low affinity, probably in a non-receptor intermediate fashion. The dissociation constant of transferrin and its receptor was calculated 9.29 nM which corresponds well to values found in the literature. In contrast to lactoferrin, the cell was capable of binding only low amounts of transferrin in a non-receptor intermediate fashion. After chemical crosslinking of lactoferrin to the cell surface, the radiolabeled lactoferrin was found in a complex of molecular mass 300 kDa. Crosslinking of transferrin resulted in a complex of much higher molecular mass. These data clearly show a binding site for lactoferrin different from the transferrin receptor. Only if competition experiments were performed with a high molar excess of both ligand proteins did a small percentage of either of the two ligands crossreact with the receptor for the other, possibly due to a structural similarity of the two glycoproteins.
Roiron
D
D
Groupe de Recherche sur les Glandes Exocrines de l'Institut National de la Santé et de la Recherche Médicale, Marseille, France.
Amouric
M
M
Marvaldi
J
J
Figarella
C
C
D000230:Adenocarcinoma / Q000378:metabolism*; D001667:Binding, Competitive; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism*; D007782:Lactoglobulins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D007782:Lactoglobulins; D011956:Receptors, Cell Surface; D014168:Transferrin; C038340:lactoferrin receptors; D007781:Lactoferrin