Detection of hemoglobin S polymerization in intact red cells by P-31 NMR

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Summary

Phosphorus-31 NMR has been used to observe 2,3-diphosphoglycerate (2,3-DPG) in intact human red cells containing hemoglobin AA or hemoglobin SS, in the oxygenated and deoxygenated state. When hemoglobin A cells are deoxygenated, the 2,3-DPG P-31 resonances broaden and shift. However, in cells containing hemoglobin S, a striking pattern of superimposed broad and narrow P-31 resonances is observed. This pattern can also be observed in fractionated cell preparations from which both irreversibly sickled cells and reticulocytes have been removed and in hemoglobin S solutions above the minimum gelling concentration. We conclude that 2,3-DPG is incorporated in the polymer phase, and that the broad and narrow peaks seen in deoxy S cell suspensions represent 2,3-DPG bound to hemoglobin S in the polymer and tetramer states respectively.

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