Characterization of the Plasmodium falciparum M17 leucyl aminopeptidase. A protease involved in amino acid regulation with potential for antimalarial drug development

Amino acids generated from the catabolism of hemoglobin by intra-erythrocytic malaria parasites are not only essential for protein synthesis but also function in maintaining an osmotically stable environment, and creating a gradient by which amino acids that are rare or not present in hemoglobin are drawn into the parasite from host serum. We have proposed that a Plasmodium falciparum M17 leucyl aminopeptidase (PfLAP) generates and regulates the internal pool of free amino acids and therefore represents a target for novel antimalarial drugs. This enzyme has been expressed in insect cells as a functional 320-kDa homo-hexamer that is optimally active at neutral or alkaline pH, is dependent on metal ions for activity, and exhibits a substrate preference for N-terminally exposed hydrophobic amino acids, particularly leucine. PfLAP is produced by all stages in the intra-erythrocytic developmental cycle of malaria but was most highly expressed by trophozoites, a stage at which hemoglobin degradation and parasite protein synthesis are elevated. The enzyme was located by immunohistochemical methods and by transfecting malaria cells with a PfLAP-green fluorescent protein construct, to the cytosolic compartment of the cell at all developmental stages, including segregated merozoites. Amino acid dipeptide analogs, such as bestatin and its derivatives, are potent inhibitors of the protease and also block the growth of P. falciparum malaria parasites in culture. This study provides a biochemical basis for the antimalarial activity of aminopeptidase inhibitors. Availability of functionally active recombinant PfLAP, coupled with a simple enzymatic readout, will aid medicinal chemistry and/or high throughput approaches for the future design/discovery of new antimalarial drugs

Metabolism of lipoproteins in Rodent malaria, Relationship between lipolysis, steatosis and increased biosynthesis of V.L.D.L.

The kinetic study of the seric free fatty acids, total lipids and hepatic triacyglycerides had led us to conclude that the biosynthesis of T.A.G.-rich lipoproteins increases during malaria. It seems that the parasite induces a lipolysis of adipose tissue in order to meet its own needs for fatty acids and that the excess of the latter taken by the liver involves an increased synthesis of the V.L.D.L. The cis-vaccenic acid has also been analysed during the evolution of parasitaemia; these variations by themselves cannot explain the extra parasitic hemolysis

Aminopeptidase of

An aminopeptidase capable of degrading synthetic substrates and short peptides has been demonstrated in Eimeria nieschulzi sporulated oocysts. Physico-chemical properties and the classic activator and inhibitory effects have been determined.The inhibitory activity of classic antimalarial drugs has been demonstrated and the study of the inhibitory mechanism by Chloroquine has been further studied.Lastly, the comparison between the aminopeptidase of Eimeria genus parasites and that of Plasmodium has been discussed