Fumarate reductase and other mitochondrial activities in Trypanosoma cruzi

Boveris, A; Hertig, Cecilia M.; Turrens, Julio F.

doi:10.1016/0166-6851(86)90121-0

Cited by 52 publications

(28 citation statements)

References 21 publications

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“…On the other hand, solublized FRD is part of the enzyme (another part of the enzyme is still located in the membrane), and its physiological position is changed, which may result in difficulty for licochalcone A in binding to the active site. Similar findings have been reported for Trypanosoma cruzi (5) and Trypanosoma brucei (24).…”

Section: Discussionsupporting

confidence: 90%

“…FRD catalyzes the reduction of fumarate to succinate, which is a key enzyme in anaerobic energy metabolism for many organisms respiring with fumarate as a terminal electron acceptor. This enzyme has been found among some bacteria such as Helicobacter pylori and Escherichia coli (16,18), among and protozoal parasites of the genera Trypanosoma (5,11,31,34), Plasmodium (14), and Leishmania (31), and in helminths (17,29). In mammalian cells, succinate is converted to fumarate by the action of SDH and then converted to malate via fumarase.…”

Section: Discussionmentioning

confidence: 99%

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Inhibition of Fumarate Reductase in Leishmania major and L. donovani by Chalcones

Chen

Zhai

Christensen

et al. 2001

Antimicrob Agents Chemother

200

130

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Our previous studies have shown that chalcones exhibit potent antileishmanial and antimalarial activities in vitro and in vivo. Preliminary studies showed that these compounds destroyed the ultrastructure of Leishmania parasite mitochondria and inhibited the respiration and the activity of mitochondrial dehydrogenases of Leishmania parasites. The present study was designed to further investigate the mechanism of action of chalcones, focusing on the parasite respiratory chain. The data show that licochalcone A inhibited the activity of fumarate reductase (FRD) in the permeabilized Leishmania major promastigote and in the parasite mitochondria, and it also inhibited solubilized FRD and a purified FRD from L. donovani. Two other chalcones, 2,4-dimethoxy-4-allyloxychalcone (24m4ac) and 2,4-dimethoxy-4-butoxychalcone (24mbc), also exhibited inhibitory effects on the activity of solubilized FRD in L. major promastigotes. Although licochalcone A inhibited the activities of succinate dehydrogenase (SDH), NADH dehydrogenase (NDH), and succinate-and NADHcytochrome c reductases in the parasite mitochondria, the 50% inhibitory concentrations (IC 50 ) of licochalcone A for these enzymes were at least 20 times higher than that for FRD. The IC 50 of licochalcone A for SDH and NDH in human peripheral blood mononuclear cells were at least 70 times higher than that for FRD. These findings indicate that FRD, one of the enzymes of the parasite respiratory chain, might be the specific target for the chalcones tested. Since FRD exists in the Leishmania parasite and does not exist in mammalian cells, it could be an excellent target for antiprotozoal drugs.Leishmaniasis is a major and increasing public health problem, particularly in Africa, Asia, and Latin America (23, 37). Some 350 million people are at risk of infection with Leishmania spp., and more than 12 million people are infected with different species of the parasite. Each year, there are 1.5 million new cases, and 500,000 of these are visceral leishmaniasis, which is nearly always fatal if left untreated (23). Treatment of leishmaniasis is unsatisfactory in that the existing drugs require repeated parenteral administration, and none of them are effective in all cases or are totally free of side effects (1, 26, 37). Furthermore, large-scale clinical resistance to antimonials, the first-line antileishmanial drugs, has been reported recently. This resistance occurred in 5 to 70% of patients in some areas of endemicity (28, 36). There is, therefore, a great and urgent need for the development of new, effective, and safe drugs for the treatment of leishmaniasis.A number of investigations to explore potential antileishmanial drugs have been carried out during the last 2 decades (2,6,15,21,22,25,30,33,38). We have previously reported that chalcones have potent antileishmanial and antimalarial activities and might be developed into a new class of antileishmanial drugs (7-10, 39). Attempting to elucidate the antileishmanial mechanism of action of the chalcones, we have previously fou...

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Inhibition of Fumarate Reductase in Leishmania major and L. donovani by Chalcones

Chen

Zhai

Christensen

et al. 2001

Antimicrob Agents Chemother

200

130

View full text Add to dashboard Cite

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“…The identification of FRDm1 as the mitochondrial NADH-FRD seems to bring the long standing debate regarding the existence and the subcellular localization of FRD enzymes in trypanosomatids (27)(28)(29) to an end. NADH-FRD activity was also detected in mitochondrial extracts of T. cruzi (30,48) and Leishmania donovani (49). In agreement with these biochemical data, the recently sequenced L. major and T. cruzi genomes contain one and two FRD genes with putative mitochondrial targeting motifs, respectively, in addition to a glycosomal FRD gene (data not shown).…”

Section: Discussionsupporting

confidence: 74%

“…The glycosomal and mitochondrial compartments and the tricarboxylic acid cycle are indicated. AA, amino acid; 1,3BPGA, 1,3-bisphosphoglycerate; C, cytochrome c; Cit, citrate; CoASH, coenzyme A; F-6-P, fructose 6-phosphate; FBP, fructose 1,6-bisphosphate; G-3-P, glyceraldehyde 3-phosphate; G-6-P, glucose 6-phosphate; GLU, glutamate; Gly-3-P, glycerol 3-phosphate; IsoCit, isocitrate; 2Ket, 2- T. cruzi (6,30,31). However, it was suggested that this activity might correspond to a reversal of the reaction catalyzed by the mitochondrial SDH (27).…”

mentioning

confidence: 99%

A Mitochondrial NADH-dependent Fumarate Reductase Involved in the Production of Succinate Excreted by Procyclic Trypanosoma brucei

Coustou

Besteiro

Rivière

et al. 2005

Journal of Biological Chemistry

115

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Trypanosoma brucei is a parasitic protist responsible for sleeping sickness in humans. The procyclic stage of T. brucei expresses a soluble NADH-dependent fumarate reductase (FRDg) in the peroxisome-like organelles called glycosomes. This enzyme is responsible for the production of about 70% of the excreted succinate, the major end product of glucose metabolism in this form of the parasite. Here we functionally characterize a new gene encoding FRD (FRDm1) expressed in the procyclic stage. FRDm1 is a mitochondrial protein, as evidenced by immunolocalization, fractionation of digitonin-permeabilized cells, and expression of EGFP-tagged FRDm1 in the parasite. RNA interference was used to deplete FRDm1, FRDg, or both together. The analysis of the resulting mutant cell lines showed that FRDm1 is responsible for 30% of the cellular NADH-FRD activity, which solves a long standing debate regarding the existence of a mitochondrial FRD in trypanosomatids. FRDg and FRDm1 together account for the total NADH-FRD activity in procyclics, because no activity was measured in the double mutant lacking expression of both proteins. Analysis of the end products of 13 C-enriched glucose excreted by these mutant cell lines showed that FRDm1 contributes to the production of between 14 and 44% of the succinate excreted by the wild type cells. In addition, depletion of one or both FRD enzymes results in up to 2-fold reduction of the rate of glucose consumption. We propose that FRDm1 is involved in the maintenance of the redox balance in the mitochondrion, as proposed for the ancestral soluble FRD presumably present in primitive anaerobic cells.Fumarate reductases (FRDs) 1 catalyze the reduction of fumarate to succinate and can be divided into two classes of enzymes: those belonging to a multimeric complex associated with the respiratory chain and transferring electrons from a quinol to fumarate and the soluble enzymes, which transfer electrons from a noncovalently bound cofactor (NADH or FADH 2 /FMNH 2 ) to fumarate. Most of the FRDs characterized so far belong to the first class (1). These FRDs are structurally similar to succinate dehydrogenases (SDH), complex II of the respiratory chain. The Shewanella putrefaciens FRD is functionally equivalent to the membrane-bound enzymes by accepting/transferring electrons from/to the respiratory chain but is a soluble enzyme lacking a membrane anchor (2). To date, only two examples of soluble FRDs not linked to the respiratory chain (second class) have been described. The yeast Saccharomyces cerevisiae expresses two soluble FRDs (cytosolic and promitochondrial) that use FADH 2 /FMNH 2 as an electron donor (3, 4), and the African trypanosome Trypanosoma brucei expresses a soluble NADH-dependent FRD located in the peroxisome-like organelle, called glycosome (5, 6). A phylogenetic analysis of FRDs and SDHs showed that the membrane-bound enzymes form a monophyletic group distantly related to the soluble enzymes, including the S. putrefaciens FRD (5). In 1980, Gest (7) proposed that the membrane-bou...

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“…Nevertheless, the mechanisms by which the substitutioncontaining chalcones showed leishmanicidal and trypanocidal activities were not addressed in this work. Based on the literature, it can be predicted that chalcones could potentially inhibit the activity of fumarate reductase, succinate dehydrogenase, NADH dehydrogenase, or succinate-and NADH-cytochrome c reductases in the parasite mitochondria (2,3,7,19,20). Additional studies are in progress to address this hypothesis.…”

mentioning

confidence: 99%

Trypanocidal and Leishmanicidal Properties of Substitution-Containing Chalcones

Lunardi

Guzela

Rodrigues

et al. 2003

Antimicrob Agents Chemother

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Ten chalcones were synthesized and tested as potential leishmanicidal and trypanocidal agents. All tested compounds caused concentration-dependent inhibition of the in vitro growth of Leishmania braziliensis and Trypanosoma cruzi with no significant toxic effect towards host macrophages. Our results show that the positions of the substituents seem to be critical for their antiprotozoal activities.Among the kinetoplastid protozoa, which infect invertebrates, mammals, and plants, some species are of particular interest due to their medical importance. These include Trypanosoma cruzi (the agent of Chagas' disease), the African trypanosome responsible for sleeping sickness, and several species of Leishmania, which cause the various forms of leishmaniasis (9). The World Health Organization has identified Chagas' disease and leishmaniasis as major and increasing public health problems, particularly in Latin America (14,15,16,18). In spite of the socioeconomic importance of these tropical infectious diseases, efforts directed towards the discovery of new drugs and/or vaccines against them are underdeveloped (10, 13). In addition, most of the drugs currently in use (i) were developed several decades ago, (ii) show variable efficacy, (iii) have serious side effects, (iv) are expensive, (v) can require long-term treatment, (vi) may have low activity in immunosuppressed patients, and (vii) present and/or induce resistance in parasites (9, 10, 16). Thus, the need for the development of new, effective, cheap, and safe drugs for the treatment of leishmaniasis and Chagas' disease is very important.Chalcones, or 1,3-diaryl-2-propen-1-ones, are natural or synthetic compounds belonging to the flavonoid family. Chalcones possess a broad spectrum of biological activities, including antibacterial, anthelmintic, amoebicidal, antiulcer, antiviral, insecticidal, antiprotozoal, anticancer, cytotoxic, and immunosuppressive activities (for reviews, see references 11 and 12). The present study was designed to determine the in vitro leishmanicidal and trypanocidal activities of the 10 substitutioncontaining chalcones and to investigate the cytotoxic effects of these chalcones on mouse peritoneal macrophages in vitro.The chalcones used in the present study were synthesized in our laboratory by reaction of the appropriate aryl methyl ketone and aryl aldehyde (in a 1:1 ratio) in the presence of sodium hydroxide and ethanol. The products were then added to cooled diluted acetic acid according to the methodology previously described (8). The synthetic reaction gave substantial yields (55 to 98%) of all the chalcones, and these were characterized by 1 H nuclear magnetic resonance and infrared analyses and by microanalysis. The substitution-containing chalcones were dissolved in 0.5% Tween 80 in phosphatebuffered saline to prepare a working solution with a 0.1 M concentration before being passed through 0.22-m-pore-size Millipore filters. The structures of the chalcones are shown in Table 1.Cultures of promastigote forms of Leishmania braziliensis...

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Fumarate reductase and other mitochondrial activities in Trypanosoma cruzi

Cited by 52 publications

References 21 publications

Inhibition of Fumarate Reductase in Leishmania major and L. donovani by Chalcones

Inhibition of Fumarate Reductase in Leishmania major and L. donovani by Chalcones

A Mitochondrial NADH-dependent Fumarate Reductase Involved in the Production of Succinate Excreted by Procyclic Trypanosoma brucei

Trypanocidal and Leishmanicidal Properties of Substitution-Containing Chalcones

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