Thermal studies of the rates of the reactions of carbon dioxide in concentrated haemoglobin solutions and in red blood cells. A. The reactions catalysed by carbonic anhydrase. B. The carbamino reactions of oxygenated and deoxygenated haemoglobin.
1. The initial rates of uptake of carbon dioxide by bovine Hb and HbO(2), in solutions and in intact red cells at 25 degrees C, pH 6.8-7.4, have been measured by the continuous flow rapid calorimeter in the presence and absence of the carbonic anhydrase inhibitor acetazolamide.2. Over the time range 0-10 msec the only significant reaction, in the presence of acetazolamide, was the carbamino combination of CO(2). In the absence of acetazolamide there was a much larger heat evolution owing to the simultaneous catalysis of CO(2) to bicarbonate due to the enzyme, carbonic anhydrase. Subtraction of the former rate from the latter gives the catalysis of the CO(2) hydration per se. The mode of calculating the actual amount of chemical change from the observed heat evolution is described in detail.3. The carbonic anhydrase activity was found to vary about threefold between individual blood samples, as in previous work on dilute bovine enzyme solutions, but to be independent of the state of oxygenation of the haemoglobin. Comparison of the enzyme activity in solution and in the red cell seemed to show a discrepancy of the order of 30% in favour of the latter when approximate allowance was made for the difference of pH and Cl(-) content in the two cases. This 30% difference might be reduced if exact corrections could be applied.4. No significant difference was observed between the rates of the carbamino CO(2) reactions in concentrated Hb solutions and red cell suspensions, provided that approximate allowance was made for differences of pH in both cases.5. The rate of carbamino combination with Hb appeared to be about twice that with HbO(2) at the same pH. Possible explanations for this difference are discussed.
D000086:Acetazolamide / Q000494:pharmacology*; D000818:Animals; D001639:Bicarbonates / Q000096:biosynthesis; D002151:Calorimetry; D002245:Carbon Dioxide / Q000378:metabolism*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002417:Cattle; D004912:Erythrocytes / Q000502:physiology*; D006454:Hemoglobins; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D010100:Oxygen
D001639:Bicarbonates; D006454:Hemoglobins; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases; D000086:Acetazolamide; D010100:Oxygen