Properties, stage-dependent expression and localization of Plasmodium falciparum M1 family zinc-aminopeptidase
| Autor: | Isabelle Florent, Joseph Schrével, M. Allary |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Proteases
Erythrocytes medicine.medical_treatment Amino Acid Motifs Blotting Western Plasmodium falciparum Fluorescent Antibody Technique Aminopeptidase Aminopeptidases Gene Expression Regulation Enzymologic Food vacuole medicine Animals Microscopy Phase-Contrast Enzyme Inhibitors Protease Microscopy Confocal biology Gene Expression Regulation Developmental biology.organism_classification Blot Molecular Weight Zinc Infectious Diseases Biochemistry Cytoplasm Polyclonal antibodies biology.protein Animal Science and Zoology Parasitology Electrophoresis Polyacrylamide Gel |
| Zdroj: | Parasitology. 125(Pt 1) |
| ISSN: | 0031-1820 |
| Popis: | A Plasmodium falciparum single copy gene predicting a 122 kDa protein belonging to the M1 family of zinc-metallopeptidases was previously reported and related to erythrocytic schizont proteins of 96 (p96) and 68 (p68) kDa. By using protease inhibitors during parasite harvest and enzyme preparations, and polyclonal antibodies specific for 2 peptidic domains deduced from the gene, we identified the 120 kDa precursor and demonstrated its processing into p96 and p68. The N-terminal ends of p96 and p68 were mapped between glycine-123 and lysine-163, both proteins thus containing the catalytic domain. The purified enzyme, here named PfA-M1 (p96/p68), displayed strict aminopeptidase activity, optimal at pH 7·4, with broad substrate spectrum. Its inhibition and reactivation profiles were typical of zinc-metalloaminopeptidases. By Western blotting, PfA-M1 was detected in trophozoites, in addition to schizonts, but not in early rings. PfA-M1 was localized by indirect immunofluorescence confocal microscopy. In trophozoites, the labelling was diffuse in the parasite cytoplasm, with accumulations around the food vacuole. In schizonts, it turned progressively to a vesicle-like pattern, ending as a clear spot in released merozoites. The involvement of PfA-M1 in haemoglobin breakdown and erythrocyte reinvasion is discussed in light of the dual functions recently reported for several P. falciparum proteases. |
| Databáze: | OpenAIRE |
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