Preparation of renal cortex basal-lateral and bursh border membranes. Localization of adenylate cyclase and guanylate cyclase activities.
Luminal brush border and contraluminal basal-lateral segments of the plasma membrane from the same 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+1)-ATPase. However, the specific activity of (Na+ + K+)-ATPase in brush border membranes also 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. It is proposed that guanylate cyclase, in addition to (Na+ + K+)-ATPase, be used as an enzyme "marker" for the renal basal-lateral membrane.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002116:Calcitonin / Q000494:pharmacology; D002462:Cell Membrane / Q000166:cytology / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D004837:Epinephrine / Q000494:pharmacology; D006150:Guanine Nucleotides / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000378:metabolism*; D007672:Kidney Cortex / Q000201:enzymology*; D008297:Male; D010281:Parathyroid Hormone / Q000494:pharmacology; D011453:Prostaglandins / Q000494:pharmacology; D011817:Rabbits; D014198:Trehalase / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D006150:Guanine Nucleotides; D010281:Parathyroid Hormone; D011453:Prostaglandins; D000255:Adenosine Triphosphate; D002116:Calcitonin; D005723:gamma-Glutamyltransferase; D014198:Trehalase; D000262:Adenylyl Cyclases; D006162:Guanylate Cyclase; D004837:Epinephrine