Megasomes as the targets of leucine methyl ester in Leishmania amazonensis amastigotes.
Certain L-amino acid esters, such as L-leucine methyl ester (Leu-OMe), can kill intracellular and isolated Leishmania amazonensis amastigotes. Killing appears to involve ester trapping and hydrolysis within an acidified parasite compartment (M. Rabinovitch and S. C. Alfieri, 1987, Brazilian Journal of Medical and Biological Research 20, 665-74). We show here by acid phosphatase light microscopic cytochemistry and by ultrastructural morphometry that megasomes, lysosome-like amastigote organelles, are the putative parasite targets of Leu-OMe. This conclusion is supported by the following observations. (a) Control amastigotes displayed a string of electron-dense, acid phosphatase-positive megasomes mostly located in the cellular poles opposite the flagellar pockets. Incubation of the amastigotes with Leu-OMe resulted in concentration-dependent swelling and fusion of the organelles as well as decreased electron density of the internal contents. These changes, which preceded parasite disruption, were followed by the progressive loss of parasite viability and the release of acid phosphatase activity into the medium. (b) Incubation of the amastigotes with L-isoleucine methyl ester, a non-leishmanicidal compound, induced only moderate fusion of the megasomes. (c) Pre-incubation of the parasites with the proteinase inhibitors antipain and chymostatin, previously shown to confer protection from Leu-OMe toxicity, nearly completely prevented the morphological changes of megasomes. (d) Exposure of amastigotes to tryptophanamide (Trp-NH2), the leishmanicidal activity of which is not reduced by antipain and chymostatin, did not result in swelling and fusion of the megasomes. This last finding suggests that different mechanisms underlie the destruction of amastigotes by Trp-NH2 and Leu-OMe.(ABSTRACT TRUNCATED AT 250 WORDS)
Antoine
J C
JC
Unité d' Immunophysiologie cellulaire de l'Institut Pasteur et du C.N.R.S. (UA 1113), Département de Physiopathologie expérimentale, Paris, France.
Jouanne
C
C
Ryter
A
A
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D004136:Dinitrobenzenes / Q000494:pharmacology; D005260:Female; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D007891:Leishmania / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D007930:Leucine / Q000494:pharmacology / Q000031:analogs & derivatives; D008247:Lysosomes / Q000187:drug effects* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D011480:Protease Inhibitors / Q000494:pharmacology
D004136:Dinitrobenzenes; D011480:Protease Inhibitors; C030365:leucine methyl ester; C042432:3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine; D000135:Acid Phosphatase; D007930:Leucine