Photoaffinity labelling of Plasmodium falciparum proteins involved in phospholipid transport

Mol Biochem Parasitol. 1994 Oct;67(2):235-43. doi: 10.1016/0166-6851(94)00136-7.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / pharmacology
  • Affinity Labels*
  • Animals
  • Azides / metabolism
  • Biological Transport / drug effects
  • Cell Membrane / metabolism
  • Cross-Linking Reagents
  • Electrophoresis, Polyacrylamide Gel
  • Erythrocytes / parasitology
  • Fluorescent Dyes
  • Humans
  • Phosphatidylethanolamines / metabolism
  • Phospholipids / metabolism*
  • Plasmodium falciparum / metabolism*
  • Protozoan Proteins / metabolism*
  • Salicylates / metabolism

Substances

  • Affinity Labels
  • Azides
  • Cross-Linking Reagents
  • Fluorescent Dyes
  • Phosphatidylethanolamines
  • Phospholipids
  • Protozoan Proteins
  • Salicylates
  • 1,2-dilauroylphosphatidylethanolamine
  • N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine
  • Adenosine Triphosphate
  • 4-azidosalicylic acid N-hydroxysuccinimide ester