Human Renal Carbonic Anhydrase. Purification and Properties

Abstract

Carbonic anhydrase was isolated from fresh human donor kidneys which had been thoroughly perfused free from blood. The isolation procedure involved biospecific affinity chromatography on a sulfanilamide–agarose column and yielded one soluble form of the enzyme, which was homogenous with respect to sedimentation in the ultracentrifuge, electrophoresis, isoelectric focusing and immunodiffusion. The renal enzyme had an amino acid composition and behaved chromatographically, electrophoretically, and immunochemically like the erythrocyte form human carbonic anhydrase C, isolated by the same technique. The Kinetic behaviour of the renal enzyme was similar to that of human carbonic anhydrase C when compared by the stopped-flow pH-indicator technique. The results therefore suggest that the cytoplasmic carbonic anhydrase of the human kidney is very similar, if not identical, to the high-activity erythrocyte form of human carbonic anhydrase C.