title: Plasmodium berghei and Plasmodium chabaudi: a neutral endopeptidase in parasite extracts and plasma of infected animals.
abstract: By using a sensitive fluorometric method with Val-Leu-Gly-Arg-3-amino-9-ethylcarbazole (VLGR-AEC) as a substrate, two endopeptidase activities were identified in two fractions of Sephacryl S-200 gel filtration from soluble P. berghei and P. chabaudi extracts. Controls with normal mouse erythrocytes, with leukocytes, and with reticulocyte enriched blood and different washing procedures during the preparation of soluble P. berghei extracts showed that the MW greater than 200 kDa fraction was a contaminant from erythrocytes and exhibited an optimal pH activity of 8.2. In contrast, the fraction 130 kDa was related to P. berghei and P. chabaudi and exhibited an optimal pH activity of 7.4. The two enzyme activities were compared with eight different substrates. The parasite endopeptidase showed a strong activity with Val-Leu-Gly-Lys-AEC (VLGK-AEC) and Ser-Gly-Lys-AEC (SGK-AEC) as substrates; in contrast, the mouse host endopeptidase poorly cleaved the VLGK-AEC and did not cleave SGK-AEC. Presence of the hydrophobic benzyl group on serine reduced the hydrolizing properties of P. berghei endopeptidase: the reverse was observed with host endopeptidase. The hydrolysis of the N-polyhydroxyalcanoyl-VLGK-AEC substrate by the parasite neutral endopeptidase strongly increased with the schizogonic stage, as shown with synchronized P. chabaudi in mice. By its physiological pH and specificity the release of this enzyme in mouse plasma during the infection could be of interest in a peptidyl-drug strategy.
authors.forename: F
authors.initials: F
authors.lastname: Bernard
authors.forename: R
authors.initials: R
authors.lastname: Mayer
authors.forename: I
authors.initials: I
authors.lastname: Picard
authors.forename: A
authors.initials: A
authors.lastname: Deguercy
authors.forename: M
authors.initials: M
authors.lastname: Monsigny
authors.forename: J
authors.initials: J
authors.lastname: Schrevel
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D010450:Endopeptidases / Q000032:analysis* / Q000097:blood; D006863:Hydrogen-Ion Concentration; D008288:Malaria / Q000201:enzymology*; D051379:Mice; D008970:Molecular Weight; D010961:Plasmodium / Q000201:enzymology*; D010962:Plasmodium berghei / Q000201:enzymology*
chemical_list: D010450:Endopeptidases