Roles for two aminopeptidases in vacuolar hemoglobin catabolism in Plasmodium falciparum.
During the erythrocytic stage of its life cycle, the human malaria parasite Plasmodium falciparum catabolizes large quantities of host-cell hemoglobin in an acidic organelle, the food vacuole. A current model for the catabolism of globin-derived oligopeptides invokes peptide transport out of the food vacuole followed by hydrolysis to amino acids by cytosolic aminopeptidases. To test this model, we have examined the roles of four parasite aminopeptidases during the erythrocytic cycle. Localization of tagged aminopeptidases, coupled with biochemical analysis of enriched food vacuoles, revealed the presence of amino acid-generating pathways in the food vacuole as well as the cytosol. Based on the localization data and in vitro assays, we propose a specific role for one of the plasmodial enzymes, aminopeptidase P, in the catabolism of proline-containing peptides in both the vacuole and the cytosol. We establish an apparent requirement for three of the four aminopeptidases (including the two food vacuole enzymes) for efficient parasite proliferation. To gain insight into the impact of aminopeptidase inhibition on parasite development, we examined the effect of the presence of amino acids in the culture medium of the parasite on the toxicity of the aminopeptidase inhibitor bestatin. The ability of bestatin to block parasite replication was only slightly affected when 19 of 20 amino acids were withdrawn from the medium, indicating that exogenous amino acids cannot compensate for the loss of aminopeptidase activity. Together, these results support the development of aminopeptidase inhibitors as novel chemotherapeutics directed against malaria.
Dalal
Seema
S
Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA.
Klemba
Michael
M
D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D003593:Cytoplasm / Q000201:enzymology; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D016778:Malaria, Falciparum / Q000188:drug therapy / Q000201:enzymology*; D010963:Plasmodium falciparum / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014617:Vacuoles / Q000201:enzymology*
D000596:Amino Acids; D006454:Hemoglobins; D011480:Protease Inhibitors; D015800:Protozoan Proteins; D000626:Aminopeptidases; C022750:X-Pro aminopeptidase; D007930:Leucine; C012211:ubenimex