Calorimetric and optical characterization of sickle cell hemoglobin gelation. |
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Authors: | P D Ross J Hofrichter W A Eaton |
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Affiliation: | Laboratories of Molecular Biology and Chemical Physics National Institute of Arthritis, Metabolism and Digestive Diseases National Institutes of Health Bethesda, Md 20014, U.S.A. |
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Abstract: | We have used two techniques to characterize the gelation of deoxyhemoglobin S, a high sensitivity heat-flow calorimeter to measure the heat of gelation and a simple light-transmission method to measure the optical birefringence resulting from the alignment of deoxyhemoglobin S fibers in the gel. A theory for the interpretation of the birefringence measurements is presented. We combine the results of the calorimetric and optical measurements with those of sedimentation experiments to obtain enthalpy changes for gelation. The enthalpy change obtained from scanning and isothermal calorimetric measurements (0.25 m-potassium phosphate, 0.05 m-sodium dithionite, pH 6.9) varies from 4000 to 2200 cal mol−1 hemoglobin between 16 and 25 °C. There is a large apparent heat capacity change of −130 to −190 cal deg.−1 mol−1. The apparent enthalpy change estimated from solubility measurements and birefringence melting experiments is 2200 ± 500 cal mol−1 in qualitative agreement with the calorimetric results. Analysis of the time dependence of the calorimetric and optical progress curves at 20 °C leads to a rough estimate of 1800 to 4000 and −800 to 1500 cal mol−1 hemoglobin for the enthalpies of polymerization and alignment of fibers, respectively. The small magnitude of the observed enthalpy change is in accord with the view that no large conformational change takes place in the deoxyhemoglobin S molecule upon gelation. |
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