Further characterization of an early expressed glycoprotein of the rabbit small intestinal brush border. Its interaction with some hydrolases.
Mouse 23 B 921 monoclonal antibody recognized a rabbit brush-border antigen with an apparent molecular mass 140 kDa (140-kDa Ag) which, unlike most hydrolases, is expressed in the poorly differentiated crypt cells of the small intestine. Immunoelectron microscopy of brush-border vesicles showed that the 23 B 921 bound to an epitope localized on the outside of the membrane. As is the case with hydrolases the external domain of the 140-kDa Ag constitutes the main part of the molecule, which can be released by papain treatment of brush-border vesicles. The presence of a small hydrophobic domain, anchoring the molecule into the membrane and responsible for its amphipatic character, was shown by its affinity for Triton-X114 micelles. The topological organization in the membrane of 140-kDa Ag seemed to be identical to that of hydrolases. Unlike hydrolases, however, the native structure of the antigen was found from its sensitivity to proteolysis to be very dependent on its integration into the lipid bilayer. Nevertheless, detergent-extracted 140-kDa Ag can be purified by immunoaffinity chromatography although it cannot be stocked for more than 48 h even in the presence of protease inhibitors. The carbohydrate moiety of 140-kDa Ag, bearing the human blood group A determinant, amounts to 20% of the total molecular mass. The existence of some privileged relationship was established between 140-kDa Ag and hydrolases: in the membrane, hydrolases protect the 140-kDa Ag from papain action; after detergent extraction, 140-kDa Ag is strongly associated with several hydrolases particularly disaccharidases.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000302:isolation & purification*; D002241:Carbohydrates / Q000032:analysis; D002454:Cell Differentiation; D004186:Disaccharidases / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification*; D006867:Hydrolases / Q000302:isolation & purification*; D007413:Intestinal Mucosa / Q000032:analysis*; D008565:Membrane Proteins / Q000302:isolation & purification*; D008871:Microvilli / Q000032:analysis; D011817:Rabbits
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D002241:Carbohydrates; D006023:Glycoproteins; D008565:Membrane Proteins; D006867:Hydrolases; D004186:Disaccharidases