Phospholipid metabolism of serine in Plasmodium-infected erythrocytes involves phosphatidylserine and direct serine decarboxylation.
Erythrocytes infected with Plasmodium falciparum or Plasmodium knowlesi efficiently incorporated radioactive serine into phosphatidylserine (PtdSer), phosphatidylethanolamine (PtdEtn) and phosphatidylcholine (PtdCho). Serine was also metabolized into ethanolamine (Etn) and phosphorylethanolamine (P-Etn) via direct serine decarboxylation; this is a major phenomenon since together these metabolites represent 60% of total radioactive water-soluble metabolites. They were identified by reverse-phase HPLC and two TLC-type analyses and confirmed by alkaline phosphatase treatment, which depleted the radioactive P-Etn peak completely with a concomitant increase in that of Etn. In the presence of 5 microM labelled serine, radioactivity appeared in Etn and P-Etn after a 25 min lag period, and isotopic equilibrium was reached at 40 and 95 min respectively. There was a similar lag period for PtdEtn formation, which accumulated steadily for at least 180 min. Incorporation of serine into phospholipids and water-soluble metabolites increased in the presence of up to 500 microM external serine. An apparent plateau was then reached for all metabolites except intracellular serine and Etn. Exogenous Etn (at 20 microM) induced a concomitant dramatic decrease in serine incorporation into P-Etn and all phospholipids, but not into Etn. Increasing exogenous serine to 100 microM decreased the incorporation of radioactive Etn into PtdEtn by only 30%, and the PtdCho level was not affected. 2-Hydroxyethylhydrazine significantly decreased serine incorporation into P-Etn and PtdEtn, whereas Etn was accumulated. No concomitant inhibition of PtdSer or PtdCho labelling from serine occurred, even when PtdEtn formation was decreased by 95%. This indicates that the PtdEtn pool derived from direct serine decarboxylation differed from that derived from PtdSer decarboxylation, and the latter appeared to be preferentially used for PtdCho biosynthesis. Hydroxylamine also inhibited phosphorylation of serine-derived Etn but not that of exogenous Etn. The rate of PtdSer synthesis from 10 microM L-serine was 3.1+/-0.5 and 2.95+/-1.3 nmol/5 h per 10(10) infected cells, whereas L-serine decarboxylation accounted for 7.1+/-1.5 and 9.9+/-3 nmol/5 h per 10(10) infected cells for P. falciparum and P. knowlesi respectively (means+/-S.E.M.). The serine decarboxylating reaction was not detected in other higher eukaryotic cells such as mouse fibroblasts and human lymphocytes. Finally, these results also indicate compartmentalization of phospholipid metabolism in Plasmodium-infected erythrocytes.
Elabbadi
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UMR CNRS 5539, Département Biologie Santé, Université Montpellier II case 107, Place E. Bataillon, 34095 Montpellier Cedex 5, France.
Ancelin
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Vial
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HJ
D000818:Animals; D002262:Carboxy-Lyases / Q000097:blood*; D002451:Cell Compartmentation; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003653:Decarboxylation; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000469:parasitology*; D019856:Ethanolamine; D004983:Ethanolamines / Q000097:blood; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology; D008288:Malaria / Q000097:blood*; D008563:Membrane Lipids / Q000096:biosynthesis / Q000097:blood*; D051379:Mice; D010713:Phosphatidylcholines / Q000096:biosynthesis / Q000097:blood; D010714:Phosphatidylethanolamines / Q000096:biosynthesis / Q000097:blood; D010718:Phosphatidylserines / Q000096:biosynthesis / Q000097:blood; D010743:Phospholipids / Q000096:biosynthesis* / Q000097:blood; D010963:Plasmodium falciparum / Q000378:metabolism*; D016790:Plasmodium knowlesi / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*
D004983:Ethanolamines; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D012694:Serine; D019856:Ethanolamine; C005448:phosphorylethanolamine; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase