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Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes

Previously, we reported the development of novel small molecules that are potent inhibitors of the glycolytic enzyme phosphofructokinase (PFK) of Trypanosoma brucei and related protists responsible for serious diseases in humans and domestic animals. Cultured bloodstream-form trypanosomes, which are...

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Autores principales: Nare, Zandile, Moses, Tessa, Burgess, Karl, Schnaufer, Achim, Walkinshaw, Malcolm D., Michels, Paul A. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9971811/
https://www.ncbi.nlm.nih.gov/pubmed/36864883
http://dx.doi.org/10.3389/fcimb.2023.1129791
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author Nare, Zandile
Moses, Tessa
Burgess, Karl
Schnaufer, Achim
Walkinshaw, Malcolm D.
Michels, Paul A. M.
author_facet Nare, Zandile
Moses, Tessa
Burgess, Karl
Schnaufer, Achim
Walkinshaw, Malcolm D.
Michels, Paul A. M.
author_sort Nare, Zandile
collection PubMed
description Previously, we reported the development of novel small molecules that are potent inhibitors of the glycolytic enzyme phosphofructokinase (PFK) of Trypanosoma brucei and related protists responsible for serious diseases in humans and domestic animals. Cultured bloodstream-form trypanosomes, which are fully reliant on glycolysis for their ATP production, are rapidly killed at submicromolar concentrations of these compounds, which have no effect on the activity of human PFKs and human cells. Single-day oral dosing cures stage 1 human trypanosomiasis in an animal model. Here we analyze changes in the metabolome of cultured trypanosomes during the first hour after addition of a selected PFK inhibitor, CTCB405. The ATP level of T. brucei drops quickly followed by a partial increase. Already within the first five minutes after dosing, an increase is observed in the amount of fructose 6-phosphate, the metabolite just upstream of the PFK reaction, while intracellular levels of the downstream glycolytic metabolites phosphoenolpyruvate and pyruvate show an increase and decrease, respectively. Intriguingly, a decrease in the level of O-acetylcarnitine and an increase in the amount of L-carnitine were observed. Likely explanations for these metabolomic changes are provided based on existing knowledge of the trypanosome’s compartmentalized metabolic network and kinetic properties of its enzymes. Other major changes in the metabolome concerned glycerophospholipids, however, there was no consistent pattern of increase or decrease upon treatment. CTCB405 treatment caused less prominent changes in the metabolome of bloodstream-form Trypanosoma congolense, a ruminant parasite. This agrees with the fact that it has a more elaborate glucose catabolic network with a considerably lower glucose consumption rate than bloodstream-form T. brucei.
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spelling pubmed-99718112023-03-01 Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes Nare, Zandile Moses, Tessa Burgess, Karl Schnaufer, Achim Walkinshaw, Malcolm D. Michels, Paul A. M. Front Cell Infect Microbiol Cellular and Infection Microbiology Previously, we reported the development of novel small molecules that are potent inhibitors of the glycolytic enzyme phosphofructokinase (PFK) of Trypanosoma brucei and related protists responsible for serious diseases in humans and domestic animals. Cultured bloodstream-form trypanosomes, which are fully reliant on glycolysis for their ATP production, are rapidly killed at submicromolar concentrations of these compounds, which have no effect on the activity of human PFKs and human cells. Single-day oral dosing cures stage 1 human trypanosomiasis in an animal model. Here we analyze changes in the metabolome of cultured trypanosomes during the first hour after addition of a selected PFK inhibitor, CTCB405. The ATP level of T. brucei drops quickly followed by a partial increase. Already within the first five minutes after dosing, an increase is observed in the amount of fructose 6-phosphate, the metabolite just upstream of the PFK reaction, while intracellular levels of the downstream glycolytic metabolites phosphoenolpyruvate and pyruvate show an increase and decrease, respectively. Intriguingly, a decrease in the level of O-acetylcarnitine and an increase in the amount of L-carnitine were observed. Likely explanations for these metabolomic changes are provided based on existing knowledge of the trypanosome’s compartmentalized metabolic network and kinetic properties of its enzymes. Other major changes in the metabolome concerned glycerophospholipids, however, there was no consistent pattern of increase or decrease upon treatment. CTCB405 treatment caused less prominent changes in the metabolome of bloodstream-form Trypanosoma congolense, a ruminant parasite. This agrees with the fact that it has a more elaborate glucose catabolic network with a considerably lower glucose consumption rate than bloodstream-form T. brucei. Frontiers Media S.A. 2023-02-14 /pmc/articles/PMC9971811/ /pubmed/36864883 http://dx.doi.org/10.3389/fcimb.2023.1129791 Text en Copyright © 2023 Nare, Moses, Burgess, Schnaufer, Walkinshaw and Michels https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Nare, Zandile
Moses, Tessa
Burgess, Karl
Schnaufer, Achim
Walkinshaw, Malcolm D.
Michels, Paul A. M.
Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title_full Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title_fullStr Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title_full_unstemmed Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title_short Metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
title_sort metabolic insights into phosphofructokinase inhibition in bloodstream-form trypanosomes
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9971811/
https://www.ncbi.nlm.nih.gov/pubmed/36864883
http://dx.doi.org/10.3389/fcimb.2023.1129791
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