Microvillar iron-binding glycoproteins isolated from the rabbit small intestine.
Rabbit intestinal microvillus membranes possess high-affinity receptors for iron whose activity reflects homeostatic changes in mucosal iron transport. To isolate and characterize these membrane components, purified microvilli were radiolabelled with 59Fe(II) and solubilized in Triton X-100. 59Fe in 105000g supernatants co-eluted with a major broad protein peak (Mr approx. 100000) on gel-permeation chromatography and was rendered diffusible by Pronase digestion but not mild periodate degradation. Fluorescence studies with castor-bean lectin conjugates showed specific binding of this affinity probe exclusively to brush-border membranes in the intestinal epithelium. Affinity chromatography of solubilized membrane proteins showed binding to columns of immobilized lectin. Elution with D(+)-galactose released glycoprotein-bound 59Fe purified up to sevenfold over initial membrane extracts. The lectin bound up to 82% of protein-bound 59Fe. In contrast polyspecific antisera raised against rabbit microvilli in guinea-pigs precipitated less than 10% of solubilized radioactivity. Significantly more protein-bound 59Fe in detergent extracts of microvilli purified from bled animals interacted specifically with the lectin, suggesting that membrane glycoprotein receptors are involved in the homeostatic control of intestinal iron transport.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity; D005779:Immunodiffusion; D007421:Intestine, Small / Q000032:analysis*; D033862:Iron-Binding Proteins; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000032:analysis; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011402:Pronase; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D012276:Ricin / Q000276:immunology; D033863:Transferrin-Binding Proteins
D000906:Antibodies; D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D033863:Transferrin-Binding Proteins; C032193:glycoprotein receptor GPIb-IX; D012276:Ricin; D011402:Pronase