Receptor-mediated binding of milk lactoferrin to nursing piglet enterocytes: a model for studies on absorption of lactoferrin-bound iron.
Lactoferrin, an iron-binding glycoprotein that is abundant in milk of some species, has been suggested to play a key role in the absorption of iron in human infants. This hypothesis is based on the dominant role of lactoferrin as an iron-binding component in human milk and on the occurrence of lactoferrin receptors in brush-border membranes in infants' intestines. The piglet may be a useful model to evaluate the biological activity of lactoferrin because we have previously found the presence of a lactoferrin receptor in brush-border membranes from suckling piglets. In this study, viable enterocytes were isolated from 6- to 20-day-old suckling piglets. Binding studies were performed at 4 degrees C using 125I-labeled porcine lactoferrin. Scatchard analysis of equilibrium binding data showed an apparent binding constant (Kd) of 2 x 10(-6) M (SD = 0.6 x 10(-6)). This affinity is in close agreement with previous results obtained using isolated brush-border membrane vesicles. Bovine lactoferrin inhibited the binding of porcine lactoferrin. Porcine transferrin, however, did not affect porcine lactoferrin binding significantly. Thus, lactoferrin binding is highly specific. When enterocytes were incubated with 125I-labeled lactoferrin at 37 degrees C, the amount of cell-associated radioactivity exceeded the surface binding capacity of the cells by almost fivefold. This finding agrees with the continuous binding and subsequent internalization of 125I-labeled lactoferrin. The isolated piglet enterocyte seems to provide a useful model for further studies of the mechanism of receptor-mediated absorption of lactoferrin.
Gíslason
J
J
Department of Nutrition, University of California, Davis 95616-8669, USA.
Douglas
G C
GC
Hutchens
T W
TW
Lönnerdal
B
B
D000818:Animals; D000833:Animals, Suckling; D001667:Binding, Competitive; D002417:Cattle; D002470:Cell Survival; D004386:Duodenum / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D066298:In Vitro Techniques; D007408:Intestinal Absorption; D007457:Iodine Radioisotopes; D007501:Iron / Q000378:metabolism / Q000493:pharmacokinetics*; D007781:Lactoferrin / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D008871:Microvilli / Q000737:chemistry / Q000648:ultrastructure; D008892:Milk / Q000737:chemistry*; D008954:Models, Biological; D011956:Receptors, Cell Surface / Q000032:analysis / Q000502:physiology*; D013045:Species Specificity; D013552:Swine; D014168:Transferrin / Q000378:metabolism / Q000494:pharmacology
D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D014168:Transferrin; C038340:lactoferrin receptors; D007501:Iron; D007781:Lactoferrin