Resolution of apical and basal-lateral membrane populations from rat exorbital gland

Abstract

We performed analytical fractionation studies with the goal of isolating basal-lateral and apical membrane vesicles from epithelial cells of rat exorbital lacrimal gland. A density region designated window II contained elements of three physically and biochemically distinct membrane populations. These were resolved by countercurrent distribution in an aqueous polymer two-phase system. One population contained 50% of the NADPH-cytochrome c reductase activity recovered from window II and appeared to be of intracellular origin. A second population contained 70% of the recovered Na-K-ATPase, maximally enriched 42-fold with respect to the initial homogenate. This population was the major locus of alanine transport activity, roughly 85% of which was mediated by systems believed to be characteristic of epithelial cell basal-lateral membranes. It also contained portions of the alkaline phosphatase, galactosyltransferase, acid phosphatase, and NADPH-cytochrome c reductase activities. A third population accounted for 33% of the recovered alkaline phosphatase and was a secondary locus of Na-alanine transport activity, 45% of which could be attributed to systems believed to be characteristic of epithelial cell apical membranes.