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Preparation of renal cortex basal lateral and brush border membranes localization of adenylate cyclase and guanylate cyclase activities

, : Preparation of renal cortex basal lateral and brush border membranes localization of adenylate cyclase and guanylate cyclase activities. Biochimica et Biophysica Acta 466(3): 474-487

Luminal brush border and contraluminal basal-lateral segments of the plasma membrane from the same [rabbit] kidney cortex were prepared. The brush border membrane preparation was enriched in trehalase and .gamma.-glutamyltranspeptidase, whereas, the basal-lateral membrane preparation was enriched in (Na+ + K+)-ATPase. The specific activity of (Na+ + K+)-ATPase in brush border membranes increased relative to that in the crude plasma membrane fraction, suggesting that (Na+ + K+)-ATPase may be an intrinsic constituent of the renal brush border membrane in addition to being prevalent in the basal-lateral membrane. Adenylate cyclase had the same distribution pattern as (Na+ + K+)-ATPase, i.e., higher specific activity in basal-lateral membranes and present in brush border membranes. Adenylate cyclase in both membrane preparations was stimulated by parathyroid hormone, calcitonin, epinephrine, prostaglandins and 5'-guanylylimidodiphosphate. When the agonists were used in combination, enhancements were additive. In contrast to the distribution of adenylate cyclase, guanylate cyclase was found in the cytosol and in basal-lateral membranes with a maximal specific activity (NaN3 plus Triton X-100) 10-fold that in brush border membranes. ATP enhanced guanylate cyclase activity only in basal-lateral membranes. Guanylate cyclase, in addition to (Na+ + K+)-ATPase, may be used as an enzyme marker for the renal basal-lateral membrane.

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