Cloning, expression, purification, and characterization of Leishmania major dihydroorotate dehydrogenase

Feliciano, P.R.; Cordeiro, Artur T.; Costa-Filho, Antonio J.; Nonato, Maria Cristina

doi:10.1016/j.pep.2006.02.010

Cited by 39 publications

(49 citation statements)

References 19 publications

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“…Fumarate has an important role in several steps of the intermediary metabolism of trypanosomatids. In the cytosol, fumarate acts as a substrate of dihydroorotate dehydrogenase, an enzyme that participates in the de novo pyrimidine biosynthesis pathway [14]. In mitochondrion and glycosomes, fumarate may, like in procyclic T. brucei, participate in the succinic fermentation pathways acting as a substrate of the mitochondrial [37] and glycosomal [16] isoforms of fumarate reductase, respectively.…”

Section: Intracellular Localization Of the Lmfh Isoformsmentioning

confidence: 99%

“…The cytosolic isoform has been suggested to be involved in the production of fumarate that acts as a substrate for dihydroorotate dehydrogenase, an enzyme involved in the de novo pyrimidine biosynthesis pathway [13,14]. Furthermore, the cytosolic fumarate hydratase has also been described as being able to migrate from the cytosol to the nucleus where it plays a key role in DNA repair.…”

Section: Introductionmentioning

confidence: 99%

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Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Feliciano

Gupta

Dyszy

et al. 2012

International Journal of Biological Macromolecules

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Fumarate hydratases (FHs; EC 4.2.1.2) are enzymes that catalyze the reversible hydration of fumarate to S-malate. Parasitic protists that belong to the genus Leishmania and are responsible for a complex of vector-borne diseases named leishmaniases possess two genes that encode distinct putative FH enzymes. Genome sequence analysis of Leishmania major Friedlin reveals the existence of genes LmjF24.0320 and LmjF29.1960 encoding the putative enzymes LmFH-1 and LmFH-2, respectively. In the present work, the FH activity of both L. major enzymes has been confirmed. Circular dichroism studies suggest important differences in terms of secondary structure content when comparing LmFH isoforms and even larger differences when comparing them to the homologous human enzyme. CD melting experiments revealed that both LmFH isoforms are thermolabile enzymes. The catalytic efficiency under aerobic and anaerobic environments suggests that they are both highly sensitive to oxidation and damaged by oxygen. Intracellular localization studies located LmFH-1 in the mitochondrion, whereas LmFH-2 was found predominantly in the cytosol with possibly also some in glycosomes. The high degree of sequence conservation in different Leishmania species, together with the relevance of FH activity for the energy metabolism in these parasites suggest that FHs might be exploited as targets for broad-spectrum antileishmanial drugs.

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Section: Intracellular Localization Of the Lmfh Isoformsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Feliciano

Gupta

Dyszy

et al. 2012

International Journal of Biological Macromolecules

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“…Despite the fact that the enzymatic function of DHODH is conserved in all organisms, the enzyme protein structure is quite different in prokaryotic and eukaryotic organisms. The predictive enzyme from Leishmania major has been cloned expressed in Escherichia coli crystallized and the structure solved by molecular replacement technique [29,30].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the leishmanicidal and cytotoxic effects of inhibitors for microorganism metabolic pathway enzymes

Barbosa

Costa

Rocha

et al. 2015

Biomedicine & Pharmacotherapy

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“…This is the case in Babesia bovis , B. bigemina (Gero et al, 1983), Plasmodium falciparum (Krungkrai, 1995), and Toxoplasma gondii (Hortua Triana et al, 2012), where the enzyme is intimately connected to the electron transport chain to which it passes electrons directly, probably at the ubiquinone level. In contrast, in certain species of Leishmania (Gero and Coombs, 1980; Hammond and Gutteridge, 1982; Feliciano et al, 2006), Crithidia fasciculata (Pascal Jr. et al, 1983) and Trypanosoma (Gutteridge et al, 1979; Hammond and Gutteridge, 1982 and 1984; Pascal Jr. et al, 1983; Takashima et al, 2002; Annoura et al, 2005), dihydroorotate dehydrogenase is located in the cytosol, and utilizes fumarate as an electron acceptor (Takashima et al, 2002; Feliciano et al, 2006). …”

Section: Enzymes Of De Novo Pyrimidine Biosynthesismentioning

confidence: 99%

Pyrimidine metabolism in schistosomes: A comparison with other parasites and the search for potential chemotherapeutic targets

Kouni

2017

Comparative Biochemistry and Physiology Part B: Biochemistry an

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Schistosomes are responsible for the parasitic disease schistosomiasis, an acute and chronic parasitic ailment that affects more than 240 million people in 70 countries worldwide. It is the second most devastating parasitic disease after malaria. At least 200,000 deaths per year are associated with the disease. In the absence of the availability of vaccines, chemotherapy is the main stay for combating schistosomiasis. The antischistosomal arsenal is currently limited to a single drug, Praziquantel, which is quite effective with a single-day treatment and virtually no host-toxicity. Recently, however, the question of reduced activity of Praziquantel has been raised. Therefore, the search for alternative antischistosomal drugs merits the study of new approaches of chemotherapy. The rational design of a drug is usually based on biochemical and physiological differences between pathogens and host. Pyrimidine metabolism is an excellent target for such studies. Schistosomes, unlike most of the host tissues, require a very active pyrimidine metabolism for the synthesis of DNA and RNA. This is essential for the production of the enormous numbers of eggs deposited daily by the parasite to which the granulomas response precipitate the pathogenesis of schistosomiasis. Furthermore, there are sufficient differences between corresponding enzymes of pyrimidine metabolism from the host and the parasite that can be exploited to design specific inhibitors or “subversive substrates” for the parasitic enzymes. Specificities of pyrimidine transport also diverge significantly between parasites and their mammalian host. This review deals with studies on pyrimidine metabolism in schistosomes and highlights the unique characteristic of this metabolism that could constitute excellent potential targets for the design of safe and effective antischistosomal drugs. In addition, pyrimidine metabolism in schistosomes is compared with that in other parasites where studies on pyrimidine metabolism have been more elaborate, in the hope of providing leads on how to identify likely chemotherapeutic targets which have not been looked at in schistosomes.

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Cloning, expression, purification, and characterization of Leishmania major dihydroorotate dehydrogenase

Cited by 39 publications

References 19 publications

Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Evaluation of the leishmanicidal and cytotoxic effects of inhibitors for microorganism metabolic pathway enzymes

Pyrimidine metabolism in schistosomes: A comparison with other parasites and the search for potential chemotherapeutic targets

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