Abstract
The incorporation of [1-14C]palmitic or [1-14C]oleic acid into phosphatidylcholine and the effect on blood group antigen expression were examined in human erythrocytes stored at 4°C for 0-3 weeks. Blood drawn into EDTA was obtained by venepuncture from healthy volunteers. A 50% suspension of washed erythrocytes was incubated in buffer containing [1-14C]fatty acid for up to 60 min at 37°C with moderate shaking. Phosphatidylcholine was extracted and analyzed for uptake of radiolabelled fatty acid and phospholipid phosphorus content. Incorporation of [1-14C]palmitic or [1-14C]oleic acid into phosphatidylcholine was reduced during storage. The mechanism for the reduction in radiolabelled fatty acid incorporation into phosphatidylcholine was a 64% (p < 0.05) reduction in membrane phospholipase A2 activity. Although human erythrocyte membranes isolated from freshly drawn blood are capable of reacylating lysophosphatidylcholine to phosphatidylcholine, with storage, a markedly different substrate preference between palmitoyl-Coenzyme A and oleoyl-Coenzyme A was observed. Lysophosphatidylcholine acyltransferase activity assayed with oleoyl-Coenzyme A was unaltered with storage. In contrast, lysophosphatidylcholine acyltransferase activity assayed with palmitoyl-Coenzyme A was elevated 5.5-fold (p < 0.05). Despite these changes, storage of erythrocytes for up to 3 weeks did not result in altered expression of the various blood group antigens investigated. We conclude that the incorporation of palmitate and oleate into phosphatidylcholine is dramatically reduced during storage of human erythrocytes. The observed differential in vitro substrate utilization suggests that distinct acyltransferases are involved in the acylation of lysophosphatidylcholine to phosphatidylcholine in human erythrocytes.
Similar content being viewed by others
References
White DA: The phospholipid composition of mammalian tissues. In G.B. Ansell, J.N. Hawthorne, R.M.C. Dawson (eds). Form and Function of Phospholipids. Elsevier, Amsterdam, 1973, pp 441-482
Choy PC, Tran K, Hatch GM, Kroeger EA: Phospholipid metabolism in the mammalian heart. Prog Lipid Res 36: 86-101, 1997
Van den Bosch H: Phosphoglyceride metabolism. Ann Rev Biochem 43: 243-277, 1974
Lands WEM: Metabolism of glycerolipids. II. The enzymatic acylation of lysolecithin. J Biol Chem 240: 2233-2237, 1960
Lands WEM, Hart P: Metabolism of glycerolipids. VI. Specificities of acylcoenzyme A: Phospholipid acyltransferase. J Biol Chem 253: 1905-1912, 1965
Choy PC, Skrzypczak M, Lee D, Jay FT: Acyl-GPC and alkenyl/alkyl-GPC:acyl-CoA acyltransferases. Biochim Biophys Acta 1348: 124-133, 1997
Mulder E, van Deenen LLM: Metabolism of red cell lipids. 3. Pathways for phospholipid renewal. Biochim Biophys Acta 106: 348-356, 1965
Agre P, Smith BL, Hartel-Schenk S: Biochemistry of the erythrocyte Rh polypeptides: A review. Yale J Biol Med 63: 461-467, 1990
Green FA: Phospholipid requirement for Rh antigenic activity. J Biol Chem 243: 5519-5524, 1968
Hughes-Jones NC, Green EJ, Hunt VA: Loss of Rh antigen activity following the action of phospholipase A2 on red cell stroma. Vox Sang 29: 184-191, 1975
Suyama K, Goldstein J: Enzymatic evidence for differences in the placement of Rh antigens within the red cell membrane. Blood 75: 255-260, 1990
Le Petit-Thevenin J, Nobil O, Boyer J: Decreased acylation of phosphatidylcholine in diabetic rat erythrocytes. Diabetes 37: 142-146, 1988
Arthur G, Choy PC: Acylation of 1-alkenyl-glycerophosphocholine and 1-acylglycerophosphocholine in guinea pig heart. Biochem J 236: 481-487, 1986
Hatch GM, Cao SG, Angel A: Decrease in cardiac phosphatidylglycerol in streptozotocin-induced diabetic rats does not affect cardiolipin biosynthesis: Evidence for distinct pools of phosphatidylglycerol in the heart. Biochem J 306: 759-764, 1995
Chown B, Lewis M: The slanted capillary method of rhesus bloodgrouping. J Clin Path 4: 464-469, 1951
Rouser G, Siakotos AN, Fleisher S: Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots. Lipids 1: 85-86, 1966
Arvidson GAE: Structural and metabolic heterogeneity of rat liver glycerophospholipids. Eur J Biochem 4: 478-486, 1968
Tardi PG, Mukherjee JJ, Choy PC: The quantitation of long-chain acyl-CoA in mammalian tissue. Lipids 27: 65-67, 1992
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254, 1976
de Vetten MP, Agre P: The Rh polypeptide is a major fatty acid-acylated erythrocyte membrane protein. J Biol Chem 263: 18193-18196, 1988
Dise CA, Goodman DBP, Rasmussen H: Definition of the pathway for membrane phospholipid fatty acid turnover in human erythrocytes. J Lipid Res 21: 292-300, 1980
Shohet SB: Mechanisms of red cell membrane lipid renewal. In L. Bolis, J.F. Hoffman, A. Leafs (eds). Membranes and Diseases. Raven Press, New York, 1976, pp 61-74
Vomel T: Properties of ATPases and energy rich phosphates in erythrocytes of young and old individuals. Gerontology 30: 22-25, 1984
Shohet SB, Nathan DG, Karnovsky, ML: Stages in the incorporation of fatty acids into red cells. J Clin Invest 47: 1096-1108, 1968
Fyrst H, Pham DV, Lubin BH, Kypers FA: Formation of vesicles by the action of acyl-CoA:1-acyllysophosphatidylcholine acyltransferase from rat liver microsomes: Optimal solubilization conditions and analysis of lipid composition and enzyme activity. Biochemistry USA 35: 2644-2650, 1996
Man RYK, Wong T, Choy PC: Effect of lysophosphoglycerides on cardiac arrythmias. Life Sci 32: 1325-1330, 1983
Myhre BA, Demaniew S, Nelson EJ: Preservation of red cell antigens during storage of blood with different anticoagulants. Transfusion 24: 499-501, 1984
Longster GH, Tovey LA, Barnes Y, Rumsby MG: Changes in the agglutinability of red cells and the inhibition of specific agglutination by plasma from human blood taken into ACD and stored at 4°C. Vox Sang 34: 193-199, 1978
Malyska H, Kleeman JE, Masouredis SP, Victoria EJ: Effects on blood group antigens from storage at low ionic strength in the presence of neomycin. Vox Sang 44: 375-384, 1983
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rusnak, A., Coghlan, G., Zelinski, T. et al. Incorporation of fatty acids into phosphatidylcholine is reduced during storage of human erythrocytes: Evidence for distinct lysophosphatidylcholine acyltransferases. Mol Cell Biochem 213, 137–143 (2000). https://doi.org/10.1023/A:1007128501636
Issue Date:
DOI: https://doi.org/10.1023/A:1007128501636