A simple isolation method for basal-lateral plasma membranes from rat kidney cortex

Membr Biochem. 1981;4(1):49-61. doi: 10.3109/09687688109065422.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism
  • Animals
  • Cell Fractionation / methods
  • Cell Membrane / enzymology
  • Cell Membrane / ultrastructure*
  • Centrifugation, Density Gradient / methods
  • Kidney Cortex / ultrastructure*
  • Kinetics
  • Male
  • Rats
  • Sodium-Potassium-Exchanging ATPase / metabolism

Substances

  • Adenylyl Cyclases
  • Sodium-Potassium-Exchanging ATPase