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Document blocked by mass action through the addition of glycerol (Figure 2). As a result trypanosomes were lysed within five minutes [44,45]. Treatment of infected rodents with the SHAM/glycerol mixture proved to be effective to clear the blood of the animals from T. brucei [46], although only with sublethal doses was permanent aparasitemia obtained [47]. If glycolysis could be blocked selectively, i.e., without affecting the equivalent enzymes of the host, one might have a promising therapy against trypanosomiasis. D. Beyond Enzyme Inhibition: Protein Routing as a Target In trypanosomes, seven enzymes involved in glycolysis, from hexokinase to phosphoglycerate kinase, are sequestered in specialized organelles, called glycosomes [48]. These microbodies are probably evolutionary relics of an endosymbiont [49] but are devoid of genetic material encoding for the glycosomal enzymes. Instead, these enzymes are encoded in the nucleus and are post-translationally imported into the glycosome. Since the import process likely involves unfolding one might envision blocking import by stabilizing the folded http://legacy.netlibrary.com/nlreader/nlReader.dll?bookid=12640&filename=Page_369.html (2 of 2) [4/5/2004 5:32:27 PM]
Document Figure 2 Glycolysis in bloodstream-form trypanosomes. All net energy production takes place in the cytosol as the result of ATP formation by pyruvate kinase. However, the majority of glycolytic enzymes are sequestered into a specialized organelle, the glycosome. There the net ATP synthesis is zero irrespective of the presence of oxygen. Under aerobic conditions the NADH produced by glyceraldehyde-3-phosphate dehydrogenase is reoxidized via the G-3-P/DHAP shuttle, which couples glycolysis to a mitochondrial glycerophosphate oxidase. Under anaerobic conditions or when the oxidase is blocked by SHAM, equimolar amounts of glycerol and 3-phosphoglycerate are formed. However, the addition of an excess of glycerol to the cytosol prevents the reoxidation of NADH. As a result trypanosomes treated with a mixture of SHAM and glycerol die in a matter of minutes. http://legacy.netlibrary.com/nlreader/nlReader.dll?bookid=12640&filename=Page_370.html (1 of 2) [4/5/2004 5:33:24 PM] Page 370
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