Energy Metabolism as an Antimalarial Drug Target

Jortzik, Esther; Becker, Katja

doi:10.1002/9783527633883.ch5

Cited by 3 publications

(2 citation statements)

References 92 publications

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“…In the asexual blood stages, it was shown, using isotopic labeling ( 2 ) and extracellular flux analysis ( 3 ), that the parasites rely mainly on glycolysis for adenosine triphosphate (ATP) production and use oxidative phosphorylation to only a minimal extent. Due to this limited metabolic flexibility, glycolytic enzymes ( 4 ), as well as upstream processes such as glucose transporters ( 5 ), were proposed as potential drug targets. These data also further highlight the importance of ATP as the main energy source for these parasites and stimulate research into ATP metabolism.…”

Section: Introductionmentioning

confidence: 99%

Real-time measurements of ATP dynamics via ATeams in Plasmodium falciparum reveal drug-class-specific response patterns

Springer,

Heimsch,

Rahlfs

et al. 2024

Antimicrob Agents Chemother

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Malaria tropica , caused by the parasite Plasmodium falciparum ( P. falciparum ), remains one of the greatest public health burdens for humankind. Due to its pivotal role in parasite survival, the energy metabolism of P. falciparum is an interesting target for drug design. To this end, analysis of the central metabolite adenosine triphosphate (ATP) is of great interest. So far, only cell-disruptive or intensiometric ATP assays have been available in this system, with various drawbacks for mechanistic interpretation and partly inconsistent results. To address this, we have established fluorescent probes, based on Förster resonance energy transfer (FRET) and known as ATeam, for use in blood-stage parasites. ATeams are capable of measuring MgATP 2− levels in a ratiometric manner, thereby facilitating in cellulo measurements of ATP dynamics in real-time using fluorescence microscopy and plate reader detection and overcoming many of the obstacles of established ATP analysis methods. Additionally, we established a superfolder variant of the ratiometric pH sensor pHluorin (sfpHluorin) in P. falciparum to monitor pH homeostasis and control for pH fluctuations, which may affect ATeam measurements. We characterized recombinant ATeam and sfpHluorin protein in vitro and stably integrated the sensors into the genome of the P. falciparum NF54 attB cell line. Using these new tools, we found distinct sensor response patterns caused by several different drug classes. Arylamino alcohols increased and redox cyclers decreased ATP; doxycycline caused first-cycle cytosol alkalization; and 4-aminoquinolines caused aberrant proteolysis. Our results open up a completely new perspective on drugs’ mode of action, with possible implications for target identification and drug development.

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Section: Introductionmentioning

confidence: 99%

Real-time measurements of ATP dynamics via ATeams in Plasmodium falciparum reveal drug-class-specific response patterns

Springer,

Heimsch,

Rahlfs

et al. 2024

Antimicrob Agents Chemother

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“…2013) and extracellular flux analysis (Sakata-Kato and Wirth 2016), that the parasites rely mainly on glycolysis for adenosine triphosphate (ATP) production and use oxidative phosphorylation (OXPHOS) to only a minimal extent. Due to this limited metabolic flexibility, glycolytic enzymes (Jortzik and Becker 2011), as well as upstream processes such as glucose transporters (Jiang 2022) were proposed as potential drug targets. These data also further highlight the importance of ATP as the main energy source for these parasites, and stimulate research into ATP metabolism.…”

Section: Introductionmentioning

confidence: 99%

Real-time measurements of ATP dynamics via ATeams inPlasmodium falciparumreveal drug-class-specific response patterns

Springer,

Heimsch,

Rahlfs

et al. 2023

Preprint

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Malaria tropica, caused by the parasitePlasmodium falciparum(P. falciparum) remains one of the greatest public health burdens for humankind. Due to its pivotal role in parasite survival, the energy metabolism ofP. falciparumis an interesting target for drug design. To this end, analysis of the central metabolite ATP is of great interest. So far, only cell disruptive or intensiometric ATP assays have been available in this system, with various drawbacks for mechanistic interpretation, and partly inconsistent results. To address this, we have established fluorescent probes, based on FRET and known as ATeam, for use in blood stage parasites. ATeams are capable of measuring MgATP2-levels in a ratiometric manner, thereby facilitatingin cellulomeasurements of ATP dynamics in real-time using fluorescence microscopy and plate reader detection, and overcoming many of the obstacles of established ATP analysis methods. Additionally, we established a superfolder variant of the ratiometric pH sensor pHluorin (sfpHluorin) inP. falciparumto monitor pH homeostasis and control for pH fluctuations, which may affect ATeam measurements. We characterized recombinant ATeam and sfpHluorin proteinin vitroand stably integrated the sensors into the genome of theP. falciparumNF54attBcell line. Using these new tools, we found distinct sensor response patterns caused by several different drug classes. Arylamino alcohols increased and redox-cyclers decreased ATP, doxycycline caused first-cycle cytosol alkalization, and 4-aminoquinolines caused aberrant proteolysis. Our results open up a completely new perspective on drugs’ mode of action with possible implications for target identification and drug development.

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Molecular basis for covalent inhibition of glyceraldehyde‐3‐phosphate dehydrogenase by a 2‐phenoxy‐1,4‐naphthoquinone small molecule

et al. 2017

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has recently gained attention as an antiprotozoan and anticancer drug target. We have previously identified 2-phenoxy-1,4-naphthoquinone as an inhibitor of both Trypanosoma brucei and human GAPDH. Herein, through multiple chemical, biochemical, and biological studies, and through the design of analogs, we confirmed the formation of a covalent adduct, we clarified the inhibition mechanism, and we demonstrated antitrypanosomal, antiplasmodial, and cytotoxic activities in cell cultures. The overall results lent support to the hypothesis that 2-phenoxy-1,4-naphthoquinone binds the GAPDH catalytic cysteine covalently through a phenolate displacement mechanism. By investigating the reactivity of 2-phenoxy-1,4-naphthoquinone and its analogs with four GAPDH homologs, we showed that the covalent inhibition is not preceded by the formation of a strong non-covalent complex. However, an up to fivefold difference in inactivation rates among homologs hinted at structural or electrostatic differences of their active sites that could be exploited to further design kinetically selective inhibitors. Moreover, we preliminarily showed that 2-phenoxy-1,4-naphthoquinone displays selectivity for GAPDHs over two other cysteine-dependent enzymes, supporting its suitability as a warhead starting fragment for the design of novel inhibitors.

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Energy Metabolism as an Antimalarial Drug Target

Cited by 3 publications

References 92 publications

Real-time measurements of ATP dynamics via ATeams in Plasmodium falciparum reveal drug-class-specific response patterns

Real-time measurements of ATP dynamics via ATeams in Plasmodium falciparum reveal drug-class-specific response patterns

Real-time measurements of ATP dynamics via ATeams inPlasmodium falciparumreveal drug-class-specific response patterns

Molecular basis for covalent inhibition of glyceraldehyde‐3‐phosphate dehydrogenase by a 2‐phenoxy‐1,4‐naphthoquinone small molecule

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