Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion of Trypanosoma cruzi

Mantilla, Brian S.; Paes, Lisvane Silva; Pral, Elizabeth Mieko Furusho; Martil, Daiana Evelin; Thiemann, Otávio Henrique; Fernández‐Silva, Patricio; Bastos, E. L.; Silber, Ariel Mariano

doi:10.1074/jbc.m114.574525

Cited by 53 publications

(77 citation statements)

References 104 publications

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“…The substrate used was a racemic mixture of DL-⌬ 1 -pyrroline-5-carboxylate (DL-P5C) and its ring-open form ␥-GS, which was synthesized from peroxidation with NaIO 4 (Sigma) and purified by ionexchange chromatography as described previously (9,47). The steady-state activities for both TcP5CDH and TcP5CR were measured in cell-free homogenates from epimastigotes in exponential and stationary phases.…”

Section: Tcp5cdh and Tcp5cr Enzymatic Activity Assaysmentioning

confidence: 99%

“…Proteins were transferred to 0.2-m PVDF membranes (GE Healthcare), blocked with PBS buffer plus 0.3% Tween 20 (v/v) (PBST) supplemented with 5% skim milk powder (w/v), and probed for 16 h at 4°C against specific sera. TbP5CDH was probed with a polyclonal specific serum (1:4,000) raised against the recombinant TcP5CDH-His 6 (TrytripDB accession number Tc00.1047053510943.50) (9). The enzyme P5C-synthetase was probed with a polyclonal serum (1:3,000) produced in mouse against its recombinant isoform of T. cruzi (TrytripDB accession number TCSYLVIO_ 005298).…”

Section: Western Blot Analysismentioning

confidence: 99%

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Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes

Barisón

Rapado

Merino

et al. 2017

Journal of Biological Chemistry

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Edited by Chris WhitfieldTrypanosoma cruzi, the etiological agent of Chagas disease, is a protozoan parasite with a complex life cycle involving a triatomine insect and mammals. Throughout its life cycle, the T. cruzi parasite faces several alternating events of cell division and cell differentiation in which exponential and stationary growth phases play key biological roles. It is well accepted that arrest of the cell division in the epimastigote stage, both in the midgut of the triatomine insect and in vitro, is required for metacyclogenesis, and it has been previously shown that the parasites change the expression profile of several proteins when entering this quiescent stage. However, little is known about the metabolic changes that epimastigotes undergo before they develop into the metacyclic trypomastigote stage. We applied targeted metabolomics to measure the metabolic intermediates in the most relevant pathways for energy metabolism and oxidative imbalance in exponentially growing and stationary growth-arrested epimastigote parasites. We show for the first time that T. cruzi epimastigotes transitioning from the exponential to the stationary phase exhibit a finely tuned adaptive metabolic mechanism that enables switching from glucose to amino acid consumption, which is more abundant in the stationary phase. This metabolic plasticity appears to be crucial for survival of the T. cruzi parasite in the myriad different environmental conditions to which it is exposed during its life cycle.Cell growth, both in natural environments or in in vitro culture, usually presents in two phases: the exponential phase where cells divide at a roughly constant rate, and the stationary phase where cells slow down or stop the cell cycle and division. During the exponential phase, cells go through the well described canonical cell cycle of G 1 3 S 3 G 2 3 M. In the stationary phase, cells leave the regular cell cycle to enter a resting state known as quiescence, which is highly relevant because it corresponds to the physiological state in which most cells from both unicellular and multicellular organisms spend most of their life (1).Trypanosoma cruzi, the etiological agent of Chagas disease, is a protozoan parasite with a complex life cycle involving a triatomine insect and mammals. Throughout its life cycle, the T. cruzi parasite faces several alternated events of cell division and cell differentiation. Briefly, insects are infected during a blood meal from a mammal having non-dividing forms of the parasite, denominated trypomastigotes, present in their blood. Once in the midgut of the triatomine insect, the trypomastigotes differentiate to replicative non-infective epimastigotes, which proliferate and colonize the digestive tube. After the blood meal, nutrients are consumed by the intestinal epithelium, the parasite, and the intestinal microbiota population; therefore, the replicative epimastigotes eventually face nutrient starvation. Under this situation of metabolic stress, cell division is arrested, and the parasites adhe...

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Section: Tcp5cdh and Tcp5cr Enzymatic Activity Assaysmentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes

Barisón

Rapado

Merino

et al. 2017

Journal of Biological Chemistry

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show abstract

“…O P5C produzido é espontaneamente convertido a sua forma aberta glutamato-γ-semialdeído (GSA), coexistindo as duas formas de acordo com a constante de equilíbrio em meio aquoso (K = 2,2 x 10 7 ) (MANTILLA et al, 2015). Por sua vez, o GSA é substrato da segunda enzima do catabolismo de L-Pro, a P5C desidrogenase (P5CDH, EC: 1.5.1.12) que o oxida a L-Glu em uma reação dependente de NAD(P) + na membrana interna da mitocôndria, com o sítio catalítico provavelmente voltado à matriz em T. cruzi (MANTILLA et al, 2015).…”

Section: Metabolismo De L-prounclassified

Caracterização molecular e bioquímica das enzimas envolvidas na biossíntese de prolina em <i>Trypanosoma cruzi</i>.

Marchese¹

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“…Sabe-se que a Pro é degradada em dois passos enzimáticos a Glu: primeiro, é oxidada em Δ1-pirrolina-5-carboxilato (P5C), reação catalisada pela prolina desidrogenase (TcPRODH) e posteriormente o mesmo é convertido em Glu pela Δ1-pirrolina-5-carboxilato desidrogenase (TcP5CDH) na presença de NAD(P) + . Foi descrito que a oxidação de Pro acontece na mitocôndria, estimulando a respiração celular e sustentando a síntese de ATP através do processo de fosforilação oxidativa (Paes et al, 2013;Mantilla et al, 2015).…”

Section: T Cruzi E O Metabolismo De Aminoácidosunclassified

“…Foi observado que além de incorporada pelo parasita, His está sendo usada como fonte de energia para manter a viabilidade celular (Barison et al, 2016), em níveis similares aos demonstrados para Pro, anteriormente descrita como um aminoácido protetor ao estresse metabólico e relevante em termos de bioenergética para o parasita (Magdaleno et al, 2009;Martins et al, 2009;Paes et al, 2013;Mantilla et al, 2015).…”

Section: O Aminoácido Histidina (His) E O T Cruziunclassified

Caracterização molecular e funcional da imidazolona propionase de <i>Trypanosoma cruzi</i>: uma enzima do metabolismo de histidina.

Melo¹

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Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion of Trypanosoma cruzi

Abstract: Background:The enzyme, ⌬

Cited by 53 publications

References 104 publications

Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes

Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes

Caracterização molecular e bioquímica das enzimas envolvidas na biossíntese de prolina em <i>Trypanosoma cruzi</i>.

Caracterização molecular e funcional da imidazolona propionase de <i>Trypanosoma cruzi</i>: uma enzima do metabolismo de histidina.

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