pH dependency of potassium efflux from sickled red cells

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Abstract
Potassium efflux from deoxygenated, hemoglobin S‐containing red cells is often used as an “objective” in vitro measure of red cell sickling, particularly during tests with antisickling agents. Since varying pH is known to affect both the extent of sickling and passive K+‐flux across the red cell membrane, in opposite directions, we measured the sickling‐related K+‐efflux in sickle cell anemia (SS) and sickle cell trait (AS) red cells as a function of extracellular and intracellular pH. The sickling‐related K+‐efflux was found to show the same direction of pH dependence as normal red cells, so that as the extracellular pH was reduced below 7.6, sickling and K+‐efflux were increasingly dissociated. A similar dissociation was observed between sickling and K+‐efflux when the intracellular pH was lowered by increasing red cell organic phosphate levels. The sickling‐related K+‐efflux from osmotically shrunken AS cells (whose sickling tendency resembles that of SS cells) was similar in magnitude and pH dependency to that of the SS cells. The findings suggest that measurement of K+‐efflux may be an accurate estimate of the extent of intracellular polymerization in sickled red cells, provided that both the intracellular and extracellular pH levels are carefully controlled, and the experimental conditions produce no independent effects on K+ permeability.