Carbonic anhydrase assay: strong inhibition of the leaf enzyme by CO2 in certain buffers.
Apparent carbonic anhydrase activity in leaf extracts, measured as the rate of H+ production associated with the CO2 hydration reaction, varied by as much as 25-fold when the assay buffer was varied. Highest activities were usually recorded in barbitone buffer, with lower activities in imidazole, Tricine, Hepes, Tris, and phosphate buffers. The greatest differences were observed with the enzyme isolated from leaves of the monocotyledonous plants Zea mays (maize) and Triticum aestivum (wheat). Smaller differences were observed with carbonic anhydrase from dicotyledonous species and there was no effect on the erythrocyte enzyme. Leaf carbonic anhydrase activity measured by the mass spectrometric procedure was unaffected by varying the assay buffer. The low activity in certain buffers observed with the former assay system was found to be due to inhibition of the enzyme-catalyzed reaction by higher concentrations of CO2. Carbonic anhydrase from some sources was also strongly inhibited by certain inorganic and organic anions.
Hatch
M D
MD
Division of Plant Industry, CSIRO, Canberra, ACT, Australia.
D001462:Barbital / Q000494:pharmacology; D002021:Buffers; D002245:Carbon Dioxide / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D006531:HEPES / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000494:pharmacology; D010944:Plants / Q000187:drug effects / Q000201:enzymology*
D002021:Buffers; D007093:Imidazoles; D002245:Carbon Dioxide; D001462:Barbital; D002256:Carbonic Anhydrases; C018100:1-methylimidazole; D006531:HEPES