Affinity chromatography of membrane fragments. Separation of carbonic anhydrase-enriched and -depleted brain membranes.
Chick brain membranes were fractionated by affinity chromatography on AH-Sepharose 6MB linked to p-sulfamylbenzoic acid, an inhibitor of carbonic anhydrase. Two major fractions were obtained. One, designated unbound fraction, representing 42% of the total membrane protein, eluted freely from the affinity adsorbent. A second fraction, designated specifically bound fraction, contained 36% of the total membrane proteins. In addition, 11% of the total membranes bound non-specifically and could be eluted only by the use of shearing forces. Various lines of evidence indicated that the sulfonamide binding site of membrane-bound carbonic anhydrase, in addition to the carbonic anhydrase inhibitor immobilized to AH-Sepharose 6MB, was responsible for the observed specific binding. The specifically bound fraction was highly enriched in carbonic anhydrase while the unbound fraction was completely devoid of this enzyme activity.
D000110:Acetylcholinesterase / Q000032:analysis; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002091:Butyrylcholinesterase / Q000032:analysis; D002256:Carbonic Anhydrases / Q000032:analysis*; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods; D008297:Male; D008566:Membranes / Q000201:enzymology; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis; D013574:Synaptosomes / Q000201:enzymology
D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D002256:Carbonic Anhydrases; D000254:Sodium-Potassium-Exchanging ATPase