Specific antibodies to PAS IV, a glycoprotein of bovine milk-fat-globule membrane, bind to a similar protein in cardiac endothelial cells and epithelial cells of lung bronchioles

Abstract

We recently described the tissue distribution of PAS IV (periodic acid/Schiff-positive Band IV), a hydrophobic glycoprotein isolated from bovine milk-fat-globule membrane [Greenwalt and Mather (1985) J. Cell Biol. 100, 397-408]. By using immunofluorescence techniques, PAS IV was detected in mammary epithelial cells, the bronchiolar epithelium of lung, and the capillary endothelium of several tissues, including heart, salivary gland, pancreas, spleen and intestine. In the present paper we describe the specificity of the antibodies used for these studies. Two monoclonal antibodies, E-1 and E-3, were shown by solid-phase immunoassay and immunoaffinity chromatography to be specific for PAS IV (of M(r) 76000) in milk-fat-globule membrane and recognize a glycoprotein of slightly higher M(r) (85000) in heart. Affinity-purified rabbit antibodies to PAS IV were also shown to recognize components of M(r) 76000 and 85000 in fat-globule membrane and heart respectively, by using immunoblotting procedures after sodium dodecyl sulphate/polyacryamide-gel electrophoresis. Additionally, an immunoreactive protein in lung of M(r) 85000 was detected. Despite these differences in molecular size, the fat-globule membrane and heart forms of PAS IV were shown to be very similar by peptide-mapping techniques. The possible significance of the expression of similar forms of PAS IV in both epithelial and capillary endothelial cells is briefly discussed.