Abstract
Substitution of Val for Glu-β6 in the β-globin chain of hemoglobin (Hb) results in deoxy-sickle hemoglobin (HbS) forming fibers which cause sickle cell anemia. We have previously determined the three-dimensional structure of the HbS fiber from electron micrographs and synthesized a model depicting the contacts stabilizing the fiber. In order to test the contacts predicated by the model we are preparing site directed mutants. However high concentrations of hemoglobin (ca. 160 mg/ml) are required for polymers to form. Quantities of the order of 10-20 mg are available from most site directed mutant preparations. Increasing the phosphate buffer concentration to 1.5 M can reduce the concentration of HbS required for polymerization. However the use of high concentrations of phosphate buffer to induce polymerization has been criticized because there is little data relating the structure of fibers formed in high phosphate to the structure of fibers formed under physiological conditions.
Published Version
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