PMID: 3760672Oct 1, 1986Paper

Red cell vesiculation--a common membrane physiologic event

The Journal of Laboratory and Clinical Medicine
G M WagnerB Lubin

Abstract

Loss of red blood cell membrane material in the form of microvesicles has been noted in sickle cells, in Ca++-loaded and adenosine triphosphate (ATP)-depleted normal red blood cells; and during storage of normal red blood cells. To further understand the vesiculation process, we have studied vesicles generated by a variety of perturbations of the red blood cell membrane. Vesicles were isolated by centrifugation at 30,000 X g from plasma of heparinized pathologic blood samples (sickle cell anemia, hemoglobin H disease, hereditary spherocytosis, hereditary elliptocytosis, and protein 4.1 deficiency) incubated overnight at 4 degrees C. Vesicle formation also was induced in normal erythrocytes by ATP depletion, by heating to 49 degrees C, by incubation at pH 5.4, and by incubation in 5 mmol/L diamide. Membrane protein composition was characterized on denaturing polyacrylamide gels and by immunoblot. The vesicles all contained band 3, glycophorin A, and band 4.1. Spectrin was depleted in all vesicles. Thiol disulfide exchange chromatography revealed evidence of oxidative cross-linking of spectrin in pathologic and normal red blood cells that had undergone vesiculation. This suggests that the mechanism of vesiculation may be related ...Continue Reading

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