Changes in membrane polypeptides, polyphosphoinositides and phosphatidate in dense fractions of sickle cells

When sickle erythrocytes were fractionated on discontinuous isotonic stractan gradients the denser fractions, which were rich in irreversibly sickled cells contained less polyphosphoinositides and more phosphatidate than either lighter sickle cell fractions or normal cells. These changes could be due to activation of a polyphosphoinositide phosphodiesterase in the denser cells. Membrane polypeptide analysis of the denser fractions also showed a marked depletion of band 4.1 and a protein of molecular mass about 110 kDa but an increased amount of a 180 kDa polypeptide which might be a breakdown product of ankyrin. These biochemical alterations could be consequences of Ca2+ accumulation in the denser sickle cells and may contribute to the structural alterations which give rise to irreversibly sickled cells.