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The active transport of histidine and its role in ATP production in Trypanosoma cruzi

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Abstract

Trypanosoma cruzi, the aetiological agent of Chagas’s disease, metabolizes glucose, and after its exhaustion, degrades amino acids as energy source. Here, we investigate histidine uptake and its participation in energy metabolism. No putative genes for the histidine biosynthetic pathway have been identified in genome databases of T. cruzi, suggesting that its uptake from extracellular medium is a requirement for the viability of the parasite. From this assumption, we characterized the uptake of histidine in T. cruzi, showing that this amino acid is incorporated through a single and saturable active system. We also show that histidine can be completely oxidised to CO2. This finding, together with the fact that genes encoding the putative enzymes for the histidine - glutamate degradation pathway were annotated, led us to infer its participation in the energy metabolism of the parasite. Here, we show that His is capable of restoring cell viability after long-term starvation. We confirm that as an energy source, His provides electrons to the electron transport chain, maintaining mitochondrial inner membrane potential and O2 consumption in a very efficient manner. Additionally, ATP biosynthesis from oxidative phosphorylation was found when His was the only oxidisable metabolite present, showing that this amino acid is involved in bioenergetics and parasite persistence within its invertebrate host.

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Acknowledgments

This work was supported by grants from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grant #2013/18970-6 to AMS) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq grant #2013/18970-6 and #308351/2013-4 to AMS).

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Authors and Affiliations

  1. Laboratory of Biochemistry of Tryps - LaBTryps, Departament of Parasitology, Institute of Biomedical Sciences, University of São Paulo, Prédio Biomédicas II, Av. Lineu Prestes 1374, Sala 24, Cidade Universitária, (05508-000), São Paulo (SP), Brasil

    M. J. Barisón, F. S. Damasceno, B. S. Mantilla & A. M. Silber

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  1. M. J. Barisón
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  2. F. S. Damasceno
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  3. B. S. Mantilla
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  4. A. M. Silber
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Correspondence to A. M. Silber.

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The authors declare that they have no conflicts of interest with the contents of this article.

Author contribution statement

All authors made major intellectual contributions to this work and participated in the planning, conception and interpretation of the results. MJB, FSD and BSM conducted the experimental work. MJB and AMS wrote the article. AMS was responsible for funding the experimental work.

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Fig. S1

Effect of digitonin permeabilisation on respiratory rates. a. Oxygen consumption in exponential growing cells (LIT medium culture) stimulated with 5 mM His (gray) or 5 mM Pro (black). The dashed lines indicate the variation in oxygen concentration as a function of time (right axis). The solid lines represent O2 concentration (left axis). Dig: digitonin (50 μM); ADP (250 μM); Olig: oligomycin A (0.5 μg/ml); FCCP (0.5 μM). (JPEG 594 kb)

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Barisón, M.J., Damasceno, F.S., Mantilla, B.S. et al. The active transport of histidine and its role in ATP production in Trypanosoma cruzi . J Bioenerg Biomembr 48, 437–449 (2016). https://doi.org/10.1007/s10863-016-9665-9

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