Characterization of gastric mucosal membranes. IX. Fractionation and purification of K+-ATPase-containing vesicles by zonal centrifugation and free-flow electrophoresis technique

摘要: Abstract Methods are described for purification of a vesicular membrane fraction hog gastric mucosa using differential centrifugation, density gradient separation on zonal rotors and free-flow electrophoresis. As result is obtained enriched 40-fold in terms K+-ATPase free any other enzyme marker than K+-activated p- nitrophenyl phosphatase. the 5′-nucleotidase basal Mg2+-ATPase clearly separated from latter enzymes. Osmotic shock, Triton X-100 treatment or K+ ionophores increased activity isotonic conditions, but K + -p- phosphatase not affected by these treatments, nor ATPase presence NH4+. The results suggest that electrophoretic contains major population tight vesicles, whose permeability to rate limiting activity. It concluded site internal whereas external, hence, must be mobile across membrane.