Proteomic analysis identifies novel proteins of the Maurer's clefts, a secretory compartment delivering Plasmodium falciparum proteins to the surface of its host cell

• Published in: Molecular & cellular proteomics 2005-04, Vol.4 (4), p.582-593
• Main Authors: Vincensini, Laetitia, Richert, Sophie, Blisnick, Thierry, Van Dorsselaer, Alain, Leize-Wagner, Emmanuelle, Rabilloud, Thierry, Braun Breton, Catherine
• Format: Article
• Language: English
• Subjects: Animals; Biochemistry, Molecular Biology; Blotting, Western; Deuterium; Deuterium - metabolism; DNA Primers; DNA, Protozoan; DNA, Protozoan - chemistry; Endopeptidase K; Endopeptidase K - metabolism; Erythrocyte Membrane; Erythrocyte Membrane - chemistry; Erythrocyte Membrane - drug effects; Erythrocyte Membrane - metabolism; Erythrocyte Membrane - parasitology; Genomics; Glutathione Transferase; Glutathione Transferase - metabolism; Host-Parasite Interactions; Humans; Life Cycle Stages; Life Sciences; Malaria, Falciparum; Malaria, Falciparum - parasitology; Mass Spectrometry; Membrane Proteins; Membrane Proteins - analysis; Membrane Proteins - chemistry; Membrane Proteins - isolation & purification; Membrane Proteins - metabolism; Microscopy, Fluorescence; Models, Biological; Nucleic Acid Amplification Techniques; Octoxynol; Octoxynol - pharmacology; Peptide Fragments; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Plasmodium falciparum; Plasmodium falciparum - growth & development; Plasmodium falciparum - physiology; Polymerase Chain Reaction; Protozoan Proteins; Protozoan Proteins - analysis; Protozoan Proteins - chemistry; Protozoan Proteins - isolation & purification; Protozoan Proteins - metabolism; Recombinant Fusion Proteins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism; Reproducibility of Results
• ISSN: 1535-9476; 1535-9484; 1535-9484
• DOI: 10.1074/mcp.m400176-mcp200

A novel method was validated for the efficient distinction between malaria parasite-derived and host cell proteins in mass spectrometry analyses. This method was applied to a ghost fraction from Plasmodium falciparum-infected erythrocytes containing the red blood cell plasma membrane, the erythrocyte submembrane skeleton, and the Maurer's clefts, a Golgi-like apparatus linked to and addressing parasite proteins to the host cell surface. This method allowed the identification of 78 parasite proteins. Among these we identified seven novel proteins of the Maurer's clefts based on immunofluorescence studies and proteinase K digestion assays. The products of six contiguous genes located on chromosome 5 were identified, and the location within the Maurer's clefts was established for two of them. This suggests a clustering of genes encoding Maurer's cleft proteins. Our study sheds new light on the biological function of the Maurer's clefts, which are central to the pathogenesis and to the intraerythrocytic development of P. falciparum.