Characterization of an apically derived epithelial membrane glycoprotein from bovine milk, which is expressed in capillary endothelia in diverse tissues.
A glycoprotein (PAS IV) of apparent Mr 76,000 was purified from bovine milk-fat-globule membrane and partially characterized. PAS IV contained mannose, galactose, and sialic acid as principal sugars (approximately 5.3% total carbohydrate [wt/wt]) and existed in milk in at least four isoelectric variants. The glycoprotein appeared to be an integral membrane protein by several criteria. PAS IV was recovered in the detergent phase of Triton X-114 extracts of milk-fat-globule membrane at room temperature. When bound to membrane, PAS IV was resistant to digestion by a number of proteinases, although after solubilization with non-ionic detergents, the protein was readily degraded. Amino acid analysis of the purified protein revealed a high percentage of amino acids with nonpolar residues. The location of PAS IV was determined in bovine tissues by using immunofluorescence techniques. In mammary tissue, PAS IV was located on both the apical surfaces of secretory epithelial cells and endothelial cells of capillaries. This glycoprotein was also detected in endothelial cells of heart, liver, spleen, pancreas, salivary gland, and small intestine. In addition to mammary epithelial cells, PAS IV was also located in certain other epithelial cells, most notably the bronchiolar epithelial cells of lung. The potential usefulness of this protein as a specific marker of capillary endothelial cells in certain tissues is discussed.
D000818:Animals; D002196:Capillaries / Q000032:analysis; D002417:Cattle; D004727:Endothelium / Q000032:analysis; D004848:Epithelium / Q000032:analysis; D005260:Female; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D007158:Immunologic Techniques; D007526:Isoelectric Point; D008565:Membrane Proteins / Q000032:analysis*; D008892:Milk / Q000032:analysis*; D008894:Milk Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D008970:Molecular Weight; D014018:Tissue Distribution
D006023:Glycoproteins; D008565:Membrane Proteins; D008894:Milk Proteins