Hereditary Hemolytic Anemia Associated With Abnormal Membrane Lipids: Mechanism of Accumulation of Phosphatidyl Choline

In order to study the mechanism of theaccumulation of phosphatidyl choline(PC) in erythrocytes with abnormal erythrocyte phospholipids from patientswith a hereditary hemolytic anemia, thephospholipids of the erythrocytes werelabeled radioactively. Labeling of phosphatides was achieved by both passiveequilibration with preformed phosphatides, and active "acylase"-mediated incorporation of fatty acid (FA) in thepresence of glucose, ATP and coenzymeA. The labeled cells were then reincubated in fresh compatible sera and thecatabolism of the labeled erythrocytephospholipids was followed. In addition,total acylase capacity of erythrocytestroma was determined under optimalconditions in a system with excesslysophosphatide, FA, ATP, CoA, andMg⁺⁺. No differences in passive uptakeor release of phosphatides were found between the patients’ erythrocytes andcomparable reticulocyte-rich controls. Onthe other hand, overall active incorporation of FA into PC was abnormally increased in the patients’ erythrocytes,whereas incorporation of FA intophosphatidyl-ethanolamine (PE) was decreased. However, acylase capacity forboth lysophosphatidylcholine (LPC) andlysophosphatidylethanolamine (LPE) wasnormal in the patients’ cells. This apparent paradox could be explained by thesubsequent turnover of actively incorporated PC-FA which was found to bereduced. A brief labeling experiment designed to approximate pulse-chase conditions and to label primarily PC showed aconsiderable inhibition of the subsequenttransfer of PC-FA to PE upon reincubation in fresh serum. This transfer haspreviously been shown to be responsiblefor a significant portion of PC-FA catabolism. Reincubation in hyperlipemicsera obtained from patients with liverdisease or artificially enriched with PCdid not influence the abnormal outflow ofphosphatide-FA in actively labeled cells.The findings were consistent with theconcept that PC accumulated in thesecells because of a defect in the catabolismof actively incorporated PC-FA. This defect appeared to be in the transfer ofPC-FA to PE prior to final release fromthe cell. Passive exchange pathways andthe active anabolic acylase pathway werenot abnormal in these patients’ erythrocytes.

Submitted on March 4, 1971 Revised on May 4, 1971 Accepted on May 10, 1971