Plasmepsin II, an acidic hemoglobinase from the Plasmodium falciparum food vacuole, is active at neutral pH on the host erythrocyte membrane skeleton

Sylvain Le Bonniec, Christiane Deregnaucourt, Virginie Redeker, Ritu Banerjee, Philippe Grellier, Daniel E. Goldberg, Joseph Schrével

Research output: Contribution to journalArticle

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Abstract

Plasmepsin II, an aspartic protease from the human intraerythrocytic parasite Plasmodium falciparum, is involved in degradation of the host cell hemoglobin within the acidic food vacuole of the parasite. Previous characterization of enzymatic activities from Plasmodium soluble extracts, responsible for in vitro hydrolysis of erythrocyte spectrin, had shown that the hydrolysis process occurred at pH 5.0 and involved aspartic protease(s) cleaving mainly within the SH3 motif of the spectrin α-subunit. Therefore, we used a recombinant construct of the erythroid SH3 motif as substrate to investigate the involvement of plasmepsins in spectrin hydrolysis. Using specific anti-plasmepsin II antibodies in Western blotting experiments, plasmepsin II was detected in chromatographic fractions enriched in the parasite SH3 hydrolase activity. Involvement of plasmepsin II in hydrolysis was demonstrated by mass spectrometry identification of cleavage sites in the SH3 motif, upon hydrolysis by Plasmodium extract enzymatic activity, and by recombinant plasmepsin II. Furthermore, recombinant plasmepsin II digested native spectrin at pH 6.8, either purified or situated in erythrocyte ghosts. Additional degradation of actin and protein 4.1 from ghosts was observed. Specific antibodies were used in confocal imaging of schizont-infected erythrocytes to localize plasmepsin II in mature stages of the parasite development cycle; antibodies clearly labeled the periphery of the parasites. Taken together, these results strongly suggest that, in addition to hemoglobin degradation, plasmepsin II might be involved in cytoskeleton cleavage of infected erythrocytes.

Original languageEnglish (US)
Pages (from-to)14218-14223
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number20
DOIs
StatePublished - May 14 1999
Externally publishedYes

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Erythrocyte Membrane
Plasmodium falciparum
Vacuoles
Skeleton
Membranes
Food
Spectrin
Hydrolysis
Parasites
Plasmodium
Erythrocytes
Degradation
Antibodies
Hemoglobins
Peptide Hydrolases
Schizonts
plasmepsin II
Hydrolases
Cytoskeleton
Proteolysis

ASJC Scopus subject areas

  • Biochemistry

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Plasmepsin II, an acidic hemoglobinase from the Plasmodium falciparum food vacuole, is active at neutral pH on the host erythrocyte membrane skeleton. / Le Bonniec, Sylvain; Deregnaucourt, Christiane; Redeker, Virginie; Banerjee, Ritu; Grellier, Philippe; Goldberg, Daniel E.; Schrével, Joseph.

In: Journal of Biological Chemistry, Vol. 274, No. 20, 14.05.1999, p. 14218-14223.

Research output: Contribution to journalArticle

Le Bonniec, Sylvain ; Deregnaucourt, Christiane ; Redeker, Virginie ; Banerjee, Ritu ; Grellier, Philippe ; Goldberg, Daniel E. ; Schrével, Joseph. / Plasmepsin II, an acidic hemoglobinase from the Plasmodium falciparum food vacuole, is active at neutral pH on the host erythrocyte membrane skeleton. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 20. pp. 14218-14223.
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