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Purification and identification of a neutral endopeptidase inPlasmodium falciparum schizonts and merozoites

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Abstract

An endopeptidase specific to thePlasmodium falciparum erythrocytic schizont stage and to free merozoites was detected using the fluorogenic GlcA-Val-Leu-Gly-Lys(or Arg)-AEC substrate. The enzyme was purified by high performance liquid chromatography (HPLC); its optimal activity was around pH 7.5 and its isoelectric point was 4.4. The molecular weight of the enzyme was about 68000, as demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. The endopeptidase was strongly inhibited by thiol proteinase inhibitors, leupeptin, and antipain. The possible involvement of this neutral endopeptidase in the reinvasion process is discussed.

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Abbreviations

GlcA:

gluconoyl group

AEC:

3-amino-9-ethylcarbazole

EDTA:

ethylenediaminetetraacetic acid

PHMS:

p-hydroxymercuriphenylsulfonic acid

PMSF:

phenylmethylsulfonyl fluoride

STI:

soybean trypsin inhibitor

DMEM:

Dulbecco's modified Eagle's medium

HEPES:

4-(2-hydroxyethyl)-1-piperazine ethane sulfonic acid

PBS:

phosphate-buffered saline

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

ID:

inhibitory dose

bis-TRIS:

[bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane]

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Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire, URA CNRS no 290, Université de Poitiers, 40, avenue du Recteur Pineau, F-86022, Poitiers Cédex, France

    Philippe Grellier, François Bernard & Joseph Schrével

  2. Laboratoire de Biochimie Cellulaire et Moléculaire des Glycoconjugués, Centre de Biophysique Moléculaire, CNRS et Université d'Orléans, 1, rue Haute, F-45071, Orleans Cédex 2, France

    Isabelle Picard, Roger Mayer & Michel Monsigny

  3. Max-Planck-Institut für Biochemie, D-8033, Martinsried bei München, Germany

    Hans-G. Heidrich

Authors
  1. Philippe Grellier
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  2. Isabelle Picard
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  3. François Bernard
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  4. Roger Mayer
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  5. Hans-G. Heidrich
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  6. Michel Monsigny
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  7. Joseph Schrével
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Grellier, P., Picard, I., Bernard, F. et al. Purification and identification of a neutral endopeptidase inPlasmodium falciparum schizonts and merozoites. Parasitol Res 75, 455–460 (1989). https://doi.org/10.1007/BF00930972

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  • DOI: https://doi.org/10.1007/BF00930972

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