Particulate carbonic anhydrase in homogenates of human kidney.
About 2% of human kidney carbonic anhydrase (carbonate hydro-lyase, EC 4.2.1.1) has been found in particulate fractions. Its distribution in the particulate fractions obtained by differential centrifugation suggests that it may be concentrated in the brush border. The particulate enzyme is like red cell carbonic anhydrace C in its susceptibility to inhibition by anions. Particulate carbonic anhydrase is firmly bound to the membrane and is not released by incubation at pH 10.6 and 37 degrees C or by addition of Triton X-100 or deoxycholate. In 10% Triton X-100 at pH 11.3 and 37 degrees C, the particulate enzyme is inactivated with a half time of about 20 min, and this is at least an order of magnitude slower than the inactivation of soluble enzymes in the presence or absence of membranes. The soluble enzymes are inactivated within a few minutes at 25 degrees C in 3-4% sodium dodecyl sulfate, but the particulate enzyme is relatively stable under those conditions, and its half-time of inactivation at 14 degrees C with a detergent-protein ratio of 25 was about 24 h. Gel filtration with Ultragel AcA-44 in sodium dodecyl sulfate indicates that the membrane carbonic anhydrase has a molecular weight of less than 66 000, so its stability is not due to association with large membrane fragments or vesicles. These results suggest that the membrane enzyme may be a different isozyme than the soluble carbonic anhydrases. Although present in relatively small amounts, its localization on the membrane could give it functional significance.
D000469:Alkaline Phosphatase / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D011092:Polyethylene Glycols / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013384:Succinate Cytochrome c Oxidoreductase / Q000378:metabolism
D011092:Polyethylene Glycols; D013384:Succinate Cytochrome c Oxidoreductase; D000469:Alkaline Phosphatase; D002256:Carbonic Anhydrases