A simple isolation method for basal-lateral plasma membranes from rat kidney cortex.
Basal-lateral membranes were separated in a self-orienting Percoll (modified colloidal silica) gradient from a heavy microsomal membrane fraction by centrifugation at 48,000g for 0.5 h. The (Na+--K+)-ATPase activity as a marker enzyme for the basal-lateral plasma membrane was 20-fold enriched by this procedure. The adenylate-cyclase activity measured in the basal-lateral membrane fraction was stimulated 6-fold by parathyrin and only up to 1.5-fold by arginine-vasopressin, calcitonin, or isoproterenol. The yield of basal-lateral plasma membranes was 5 to 10 percent of the amount initially present in the homogenate. The method is also applicable to the pig kidney.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient / Q000379:methods; D007672:Kidney Cortex / Q000648:ultrastructure*; D007700:Kinetics; D008297:Male; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism
D000262:Adenylyl Cyclases; D000254:Sodium-Potassium-Exchanging ATPase