Genetic and chemical analyses reveal that trypanothione synthetase but not glutathionylspermidine synthetase is essential for Leishmania infantum

Sousa, André Filipe Simões de Carvalho de; Gomes‐Alves, Ana G.; Benítez, Diego; Comini, Marcelo A.; Flohé, Leopold; Jaeger, Timo; Passos, Joana Célia dos; Stuhlmann, Friedrich; Tomás, Ana M.; Castro, Helena Carla

doi:10.1016/j.freeradbiomed.2014.05.007

Cited by 46 publications

(53 citation statements)

References 42 publications

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“…For growth-rate determination, L. infantum promastigotes, previously synchronized by three or four daily passages at 10 6 cells/mL, were seeded at 10 6 cells/mL, and their density was examined over time in Neubauer counting chambers. Transfection of promastigotes was performed as described (45). Isolated clones were transferred from agar plates containing selective drug [17.5 mg/mL phleomycin (Sigma)] into liquid medium.…”

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Mitochondrial peroxiredoxin functions as crucial chaperone reservoir inLeishmania infantum

Teixeira

Castro

Cruz

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Cytosolic eukaryotic 2-Cys-peroxiredoxins have been widely reported to act as dual-function proteins, either detoxifying reactive oxygen species or acting as chaperones to prevent protein aggregation. Several stimuli, including peroxide-mediated sulfinic acid formation at the active site cysteine, have been proposed to trigger the chaperone activity. However, the mechanism underlying this activation and the extent to which the chaperone function is crucial under physiological conditions in vivo remained unknown. Here we demonstrate that in the vector-borne protozoan parasite Leishmania infantum, mitochondrial peroxiredoxin (Prx) exerts intrinsic ATP-independent chaperone activity, protecting a wide variety of different proteins against heat stress-mediated unfolding in vitro and in vivo. Activation of the chaperone function appears to be induced by temperature-mediated restructuring of the reduced decamers, promoting binding of unfolding client proteins in the center of Prx's ringlike structure. Client proteins are maintained in a folding-competent conformation until restoration of nonstress conditions, upon which they are released and transferred to ATP-dependent chaperones for refolding. Interference with client binding impairs parasite infectivity, providing compelling evidence for the in vivo importance of Prx's chaperone function. Our results suggest that reduced Prx provides a mitochondrial chaperone reservoir, which allows L. infantum to deal successfully with protein unfolding conditions during the transition from insect to the mammalian hosts and to generate viable parasites capable of perpetuating infection.chaperone | Leishmania | peroxiredoxin

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confidence: 99%

“…Expression of His.THR-mTXNPx in the resulting mtxnpx − /+MTS. His.THR-mTXNPx clones was confirmed by Western blot and indirect immunofluorescence according to previously described procedures (13,45). promastigotes.…”

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confidence: 99%

Mitochondrial peroxiredoxin functions as crucial chaperone reservoir inLeishmania infantum

Teixeira

Castro

Cruz

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…33 Moreover, previous studies involving paullones as anti-TryS scaffolds revealed that substitution of 4-azapaullones in position 9 or 11 with alkyl or aryl groups is detrimental for target inhibition 34 , whereas paullones harboring a methyl ethylendiamine side chain at the N(5)-position (e.g., 2) inhibited both recombinant L. infantum TryS (LiTryS) and the in vitro growth of L. infantum promastigotes (insect stage). 19,20 The most potent derivative (IC 50 0.15 lM and EC 50 12 lM), containing halogen substituents in position 3 (chloro) and 9 (bromo), produced a significant depletion of trypanothione in L. infantum, which confirmed its on-target effect. 19,20 We here report an approach to develop new paullones with potent growth inhibitory activity against the infective form of T. brucei.…”

Section: Introductionmentioning

confidence: 68%

“…[6][7][8][9][10][11][12] Details of the chemical mechanisms catalyzed by TryS have been reported. [13][14][15] TryS has been suggested as chemotherapeutic target against trypanosomatid parasites based on its demonstrated indispensability for T. brucei [16][17][18] and L. infantum 19 and the complete lack of homologue sequences in the human host. Recently, N(5)-substituted paullones were identified as inhibitors of TryS.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, N(5)-substituted paullones were identified as inhibitors of TryS. 19,20 Paullones (Chart 1) are a group of 7,12-dihydroindolo [3,2-d][1]benzazepin-6(5H)-ones which besides other biological properties [21][22][23] display inhibitory activities against mammalian protein kinases of the CMGC superfamily, e.g., cyclin-dependent kinases (CDKs) [24][25][26] and glycogen synthase kinase-3 (GSK-3). 27,28 Kenpaullone (1a), 29 the prototype of the series, is used as a standard GSK-3 inhibitor which displays selectivity versus a wide array of protein kinases (e.g., ERK1, DYRK1A, VEGF-R2, PLK1, INS-R).…”

Section: Introductionmentioning

confidence: 99%

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5-Substituted 3-chlorokenpaullone derivatives are potent inhibitors of Trypanosoma brucei bloodstream forms

Orban

Korn

Benítez

et al. 2016

Bioorganic & Medicinal Chemistry

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Recent Advancement in the Search of Innovative Antiprotozoal Agents Targeting Trypanothione Metabolism

2020

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Leishmania and Trypanosoma parasites are responsible for the challenging neglected tropical diseases leishmaniases, Chagas disease, and human African trypanosomiasis, which account for up to 40,000 deaths annually mainly in developing countries. Current chemotherapy relies on drugs with significant limitations in efficacy and safety, prompting the urgent need to explore innovative approaches to improve the drug discovery pipeline. The unique trypanothione‐based redox pathway, which is absent in human hosts, is vital for all trypanosomatids and offers valuable opportunities to guide the rational development of specific, broad‐spectrum and innovative anti‐trypanosomatid agents. Major efforts focused on the key metabolic enzymes trypanothione synthetase‐amidase and trypanothione reductase, whose inhibition should affect the entire pathway and, finally, parasite survival. Herein, we will report and comment on the most recent studies in the search for enzyme inhibitors, underlining the promising opportunities that have emerged so far to drive the exploration of future successful therapeutic approaches.

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Genetic and chemical analyses reveal that trypanothione synthetase but not glutathionylspermidine synthetase is essential for Leishmania infantum

Cited by 46 publications

References 42 publications

Mitochondrial peroxiredoxin functions as crucial chaperone reservoir inLeishmania infantum

Mitochondrial peroxiredoxin functions as crucial chaperone reservoir inLeishmania infantum

5-Substituted 3-chlorokenpaullone derivatives are potent inhibitors of Trypanosoma brucei bloodstream forms

Recent Advancement in the Search of Innovative Antiprotozoal Agents Targeting Trypanothione Metabolism

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