caa a22 4500 386400555 CHVBK 20180307112157.0 cr unu---uuuuu 161130e198906 xx s 000 0 eng 10.1017/S0031182000061424 doi S0031182000061424 pii (NATIONALLICENCE)cambridge-10.1017/S0031182000061424 Phospholipid metabolism in Plasmodium-infected erythrocytes: guidelines for further studies using radioactive precursor incorporation [Elektronische Daten] The biosynthesis of phospholipids is extensive in Plasmodium knowlesi-infected simian erythrocytes due to the synthesis of membranes by this single-cell eukaryote in a host erythrocyte devoid of any pathway for lipid biosynthesis. In the present paper, we show that the incorporation of [3H]glycerol, which reflects de novo biosynthesis, is better studied at 300 μM−1 mM than at the trace doses, since this non-physiological precursor does not modify the amount of phosphatidylcholine biosynthesis from [3H]choline. Time-course incorporation of radioactive glycerol, oleate, lysophosphatidylcholine, choline, and inositol in RPMI 1640 medium containing nutrients for lipid synthesis showed that the optimum incubation time for phospholipid studies is 60-90 min, after which radioactive incorporation slows considerably. On the other hand, studies with [14C]serine revealed that incubation for 2-3 h is necessary for isotopic labelling of phosphatidylcholine via phosphatidylserine decarboxylation and phosphatidylethanolamine N-methylation. Incorporation of the various fatty acids into individual lipids was related to the molecular species composing each of them. Studies with [14C palmitoyl] lysophosphatidylcholine showed a very fast intracellular release of radioactive fatty acids, which indicates a potent lysophospholipase activity. Taken together, these data define the indispensable conditions for an experimental system suitable for further studies. Copyright © Cambridge University Press 1989 Plasmodium knowlesi nationallicence phospholipid metabolism nationallicence radioactive precursor incorporation nationallicence lipid biosynthesis nationallicence Vial H. J. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France Ancelin M. L. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France Thuet M. J. Laboratoire de Physique Biologique, Institut de Biologie, Boulevard Henri IV, 34000 Montpellier, France Philippot J. R. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France Parasitology Cambridge University Press 98/3(1989-06), 351-357 0031-1820 98:3<351 1989 98 PAR https://doi.org/10.1017/S0031182000061424 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1017/S0031182000061424 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vial H. J. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France NATIONALLICENCE 700 1- Ancelin M. L. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France NATIONALLICENCE 700 1- Thuet M. J. Laboratoire de Physique Biologique, Institut de Biologie, Boulevard Henri IV, 34000 Montpellier, France NATIONALLICENCE 700 1- Philippot J. R. CNRS UA 530, INSERM U 58, 60 rue de Navacelles, 34090 Montpellier, France NATIONALLICENCE 773 0- Parasitology Cambridge University Press 98/3(1989-06), 351-357 0031-1820 98:3<351 1989 98 PAR CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-cambridge