Lipoic acid metabolism in Trypanosoma cruzi as putative target for chemotherapy

Vacchina, Paola; Lambruschi, Daniel Andrés; Uttaro, Antonio D.

doi:10.1016/j.exppara.2018.01.017

Cited by 9 publications

(29 citation statements)

References 21 publications

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“…Later studies failed to detect binding of Ca 2ϩ to PDP1c alone (48). The T. cruzi lipoylated PDH E2 subunit has been recently identified by Western blot analysis with a polyclonal antibody that detects lipoyl moieties (49), but the gene encoding this protein has not been identified yet, and we could not investigate whether association of TcPDP with TcPDH E2 increased the sensitivity of the enzyme to Ca 2ϩ . However, in agreement with the presence of an intrinsic Ca 2ϩbinding site (47), our results indicate that Ca 2ϩ can activate TcPDP alone although with only a 2-fold increase in activity.…”

Section: Discussionmentioning

confidence: 80%

Calcium-sensitive pyruvate dehydrogenase phosphatase is required for energy metabolism, growth, differentiation, and infectivity of Trypanosoma cruzi

Lander

Chiurillo

Bertolini

et al. 2018

Journal of Biological Chemistry

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In vertebrate cells, mitochondrial Ca 2؉ uptake by the mitochondrial calcium uniporter (MCU) leads to Ca 2؉-mediated stimulation of an intramitochondrial pyruvate dehydrogenase phosphatase (PDP). This enzyme dephosphorylates serine residues in the E1␣ subunit of pyruvate dehydrogenase (PDH), thereby activating PDH and resulting in increased ATP production. Although a phosphorylation/dephosphorylation cycle for the E1␣ subunit of PDH from nonvertebrate organisms has been described, the Ca 2؉-mediated PDP activation has not been studied. In this work, we investigated the Ca 2؉ sensitivity of two recombinant PDPs from the protozoan human parasites Trypanosoma cruzi (TcPDP) and T. brucei (TbPDP) and generated a TcPDP-KO cell line to establish TcPDP's role in cell bioenergetics and survival. Moreover, the mitochondrial localization of the TcPDP was studied by CRISPR/Cas9-mediated endogenous tagging. Our results indicate that TcPDP and TbPDP both are Ca 2؉-sensitive phosphatases. Of note, TcPDP-KO epimastigotes exhibited increased levels of phosphorylated TcPDH, slower growth and lower oxygen consumption rates than control cells, an increased AMP/ATP ratio and autophagy under starvation conditions, and reduced differentiation into infective metacyclic forms. Furthermore, TcPDP-KO trypomastigotes were impaired in infecting cultured host cells. We conclude that TcPDP is a Ca 2؉-stimulated mitochondrial phosphatase that dephosphorylates TcPDH and is required for normal growth, differentiation, infectivity, and energy metabolism in T. cruzi. Our results support the view that one of the main roles of the MCU is linked to the regulation of intramitochondrial dehydrogenases.

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Section: Discussionmentioning

confidence: 80%

Calcium-sensitive pyruvate dehydrogenase phosphatase is required for energy metabolism, growth, differentiation, and infectivity of Trypanosoma cruzi

Lander

Chiurillo

Bertolini

et al. 2018

Journal of Biological Chemistry

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“…The kinetoplastid class is a diverse group of single-celled flagellate organisms, many of which are human parasites, that commonly present a reduction in or total elimination of biochemical pathways that can be augmented or provided by their hosts [ 2 , 3 ]. Diseases ignored by parasites such as Leishmania brasiliensis and Trypanosoma cruzi (kinetoplastids) affect thousands of people worldwide, and so there is an ongoing effort to identify new chemotherapies [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

Cytotoxicity of Essential Oil Cordia verbenaceae against Leishmania brasiliensis and Trypanosoma cruzi

et al. 2021

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The species Cordia verbenacea DC (Boraginaceae), known as the whaling herb and camaradinha, is a perennial shrub species native to the Atlantic Forest. Its leaves are used in folk medicine as an anti-inflammatory, analgesic, antiulcerogenic and curative agent, in the form of teas or infusions for internal or topical use. The present study aimed to verify the cytotoxicity of the essential oil and the leishmanicidal and trypanocidal potential of C. verbenacea. The essential oil was characterized by GC-MS. The in vitro biological activity was determined by anti-Leishmania and anti-Trypanosoma assays. The cytotoxixity was determined using mammalian fibroblasts. The C. verbenacea species presented α-pinene (45.71%), β-caryophyllene (18.77%), tricyclo[2,2,1-(2.6)]heptane (12.56%) as their main compounds. The essential oil exhibited strong cytotoxicity at concentrations below 250 μg/mL (LC50 138.1 μg/mL) in mammalian fibroblasts. The potent anti-trypanosome and anti-promastigote activities occurred from the concentration of 62.5 μg/mL and was considered clinically relevant. The results also demonstrate that at low concentrations (<62.5 μg/mL), the essential oil of C. verbenacea managed to be lethal for these activities. This can be considered an indication of the power used in daily human consumption. Therefore, it can be concluded that the essential oil of C. verbenacea contains a compound with remarkable antiparasitic activities and requires further research.

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“…The addition of LA did not bypass such inhibition. T. cruzi was shown to incorporate very inefficiently small monocarboxylic acids like LA, BrO, or octanoic acid (Vacchina et al, 2018), suggesting the lack of the salvage pathway.…”

Section: Trypanosomatids Likementioning

confidence: 99%

Functional characterization of the first lipoyl‐relay pathway from a parasitic protozoan

Scattolini

Lavatelli

Vacchina

et al. 2022

Molecular Microbiology

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Lipoic acid (LA) is a sulfur‐containing cofactor covalently attached to key enzymes of central metabolism in prokaryotes and eukaryotes. LA can be acquired by scavenging, mediated by a lipoate ligase, or de novo synthesized by a pathway requiring an octanoyltransferase and a lipoate synthase. A more complex pathway, referred to as “lipoyl‐relay”, requires two additional proteins, GcvH, the glycine cleavage system H subunit, and an amidotransferase. This route was described so far in Bacillus subtilis and related Gram‐positive bacteria, Saccharomyces cerevisiae, Homo sapiens, and Caenorhabditis elegans. Using collections of S. cerevisiae and B. subtilis mutants, defective in LA metabolism, we gathered evidence that allows us to propose for the first time that lipoyl‐relay pathways are also present in parasitic protozoa. By a reverse genetic approach, we assigned octanoyltransferase and amidotransferase activity to the products of Tb927.11.9390 (TblipT) and Tb927.8.630 (TblipL) genes of Trypanosoma brucei, respectively. The B. subtilis model allowed us to identify the parasite amidotransferase as the target of lipoate analogs like 8‐bromo‐octanoic acid, explaining the complete loss of protein lipoylation and growth impairment caused by this compound in T. cruzi. This model could be instrumental for the screening of selective and more efficient chemotherapies against trypanosomiases.

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Lipoic acid metabolism in Trypanosoma cruzi as putative target for chemotherapy

Cited by 9 publications

References 21 publications

Calcium-sensitive pyruvate dehydrogenase phosphatase is required for energy metabolism, growth, differentiation, and infectivity of Trypanosoma cruzi

Calcium-sensitive pyruvate dehydrogenase phosphatase is required for energy metabolism, growth, differentiation, and infectivity of Trypanosoma cruzi

Cytotoxicity of Essential Oil Cordia verbenaceae against Leishmania brasiliensis and Trypanosoma cruzi

Functional characterization of the first lipoyl‐relay pathway from a parasitic protozoan

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