Plasmodium chabaudi chabaudi and P. falciparum : inhibition of aminopeptidase and parasite growth by bestatin and nitrobestatin

Abstract The major leucine aminopeptidase of the rodent malarial parasite Plasmodium chabaudi chabaudi was partially purified using a combination of high-pressure liquid chromatography on a size-exclusion column and affinity chromatography using the aminopeptidase-specific inhibitor bestatin as the ligand. The purified enzyme showed simple Michaelis-Menten kinetics when the fluorogenic peptide analogue leucyl-7-amino-4-methyl-courmarin served as the substrate, and it was strongly inhibited by both bestatin (Ki= 50.7 ± 21.0 nM) and nitrobestatin (Ki= 2.51 ± 0.2 nM) in a competitive manner. These inhibitors were also potent blockers of the growth of P. c. chabaudi and the human parasite P. falciparum in culture, and nitrobestatin was again the more potent. Therefore, the leucine aminopeptidase represents an important target to which novel anti-malarial agents could be directed.

[1]  M. Dixon.  The determination of enzyme inhibitor constants. , 1953, The Biochemical journal.

[2]  I. Sherman,et al.  Incorporation of 14C-amino-acids by malaria (Plasmodium lophurae) IV. In vivo utilization of host cell haemoglobin , 1970 .

[3]  U. K. Laemmli,et al.  Cleavage of structural proteins during , 1970 .

[4]  C. Lambros,et al.  Synchronization of Plasmodium falciparum erythrocytic stages in culture. , 1979, The Journal of parasitology.

[5]  P. Charet,et al.  Aminopeptidase in rodent Plasmodium , 1980 .

[6]  W. Trager,et al.  Peptidases from Plasmodium falciparum cultured in vitro. , 1982, Molecular and biochemical parasitology.

[7]  C. Slomianny,et al.  Ultrastructural localization of enzymes involved in the feeding process in Plasmodium chabaudi and Babesia hylomysci. , 1983, The Journal of protozoology.

[8]  Toxicological studies on bestatin. I. Acute toxicity test in mice, rats and dogs. , 1983, The Japanese journal of antibiotics.

[9]  D. V. Vander Jagt,et al.  Purification and characterization of an aminopeptidase from Plasmodium falciparum. , 1984, Molecular and biochemical parasitology.

[10]  D. V. Vander Jagt,et al.  Comparison of proteases from chloroquine-sensitive and chloroquine-resistant strains of Plasmodium falciparum. , 1987, Biochemical pharmacology.

[11]  P. Rosenthal,et al.  A malarial cysteine proteinase is necessary for hemoglobin degradation by Plasmodium falciparum. , 1988, The Journal of clinical investigation.

[12]  B. Chait,et al.  Hemoglobin degradation in the human malaria pathogen Plasmodium falciparum: a catabolic pathway initiated by a specific aspartic protease , 1991, The Journal of experimental medicine.

[13]  J T Palmer,et al.  Antimalarial effects of peptide inhibitors of a Plasmodium falciparum cysteine proteinase. , 1991, The Journal of clinical investigation.

[14]  D. Goldberg,et al.  The pathway of hemoglobin degradation in malaria parasites. , 1992, Parasitology today.

[15]  E. Bailly,et al.  Plasmodium falciparum: differential sensitivity in vitro to E-64 (cysteine protease inhibitor) and Pepstatin A (aspartyl protease inhibitor). , 1992, The Journal of protozoology.

[16]  J. Dalton,et al.  Plasmodium berghei and Plasmodium chabaudi chabaudi: development of simple in vitro erythrocyte invasion assays , 1992, Parasitology.

[17]  P. Rosenthal,et al.  Inhibition of a Plasmodium vinckei cysteine proteinase cures murine malaria. , 1993, The Journal of clinical investigation.

[18]  R. Piper,et al.  Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. , 1993, The American journal of tropical medicine and hygiene.

[19]  J. Dalton,et al.  An Improved Medium For Plasmodium chabaudi In Vitro Erythrocyte Invasion Assays , 1993, The Journal of eukaryotic microbiology.

[20]  A. Taylor.  Aminopeptidases: structure and function , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[21]  I. Gluzman,et al.  Molecular characterization and inhibition of a Plasmodium falciparum aspartic hemoglobinase. , 1994, The EMBO journal.

[22]  G. Curley,et al.  Aminopeptidases from Plasmodium falciparum, Plasmodium chabaudi chabaudi and Plasmodium berghei , 1994, The Journal of eukaryotic microbiology.

[23]  P. Rosenthal,et al.  Plasmodium falciparum: effects of proteinase inhibitors on globin hydrolysis by cultured malaria parasites. , 1995, Experimental parasitology.

[24]  D. Goldberg,et al.  The plasmodium digestive vacuole: metabolic headquarters and choice drug target. , 1995, Parasitology today.

[25]  P. Rosenthal,et al.  Functional expression of falcipain, a Plasmodium falciparum cysteine proteinase, supports its role as a malarial hemoglobinase , 1995, Infection and immunity.

[26]  I. Gluzman,et al.  Generation of hemoglobin peptides in the acidic digestive vacuole of Plasmodium falciparum implicates peptide transport in amino acid production. , 1997, Molecular and biochemical parasitology.