Photoaffinity labelling of Plasmodium falciparum proteins involved in phospholipid transport.
Erythrocytes infected with mature-stage malaria parasites accumulate phospholipids from exogenous sources. We show that the transport of N-(7-nitrobenzy-2-oxa-1,3-diazol-4-yl)-1,2- dipalmitoyl-sn-glycero-3-phosphatidylethanolamine (N-NBD-DPPE), from the erythrocyte membrane to the intracellular malaria parasite, is dependent upon metabolic energy. A photoreactive phospholipid analogue, N-[125I]iodo-4-azidosalicylamidyl-1, 2-dilauryl-sn-glycero-3-phosphatidylethanolamine (N-125I-ASA-DLPE), has been synthesised and used in an attempt to identify proteins involved in phospholipid trafficking in malaria-infected erythrocytes. This photoreactive probe was found to preferentially label a protein with an apparent molecular weight of 22 kDa. Photolabelling of the 22-kDa protein was enhanced upon ATP depletion of malaria-infected erythrocytes.
Berman
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Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia.
Shearing
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LN
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Jinsart
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Foley
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Tilley
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D000255:Adenosine Triphosphate / Q000494:pharmacology; D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000469:parasitology; D005456:Fluorescent Dyes; D006801:Humans; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D012459:Salicylates / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D015800:Protozoan Proteins; D012459:Salicylates; C007445:1,2-dilauroylphosphatidylethanolamine; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine; D000255:Adenosine Triphosphate; C046019:4-azidosalicylic acid N-hydroxysuccinimide ester