Glycolysis as a target for the design of new anti-trypanosome drugs

Verlinde, Christophe L. M. J.; Hannaert, Véronique; Blonski, Casimir; Willson, Michèle; Périé, Jacques; Fothergill‐Gilmore, Linda A.; Opperdoes, Frederik; Gelb, Michael H.; Hól, Wim G. J.; Michels, Paul A.M.

doi:10.1054/drup.2000.0177

Cited by 210 publications

(196 citation statements)

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“…Our results also suggest the presence of an hydrophobic pocket, similar to that proposed for LDH-C4 (Wong et al 1997), that can accommodate the nonpolar allylic or propylic group of NAOx or NPOx in the active site of T. cruzi HADH-isozyme II, which might explain the selective inhibition of this isozyme in comparison with the other T. cruzi dehydrogenases and the increasing affinity for those α-keto and α-hydroxyacid substrates containing nonpolar side carbon chains (Coronel et al 1981). Recently, the glycolytic enzymes have been suggested as a possible target for anti-trypanosomatid drugs design (Bakker et al 2000, Verlinde et al 2001, Lakhdar-Ghazal et al 2002, because glycolysis provides virtually all the energy for the bloodstream form of trypanosomatids (Michels 1988, Wang 1995, Bakker et al 2000. T. cruzi HADH-isozyme II is an important isozyme due to its participation in the energy metabolism of the parasite and therefore, it has been proposed that inhibitors of this enzyme could reduce the motility and survival of this parasite (Montamat et al 1982, Gerez de Burgos et al 1984, Nogueda et al 2001.…”

Section: Discussionmentioning

confidence: 99%

Trypanosoma cruzi: inhibition of alpha-hydroxyacid dehydrogenase isozyme II by N-allyl and N-propyl oxamates and their effects on intact epimastigotes

Chena¹,

Elizondo-Jiménez²,

Rodrı́guez-Páez³

et al. 2004

Mem. Inst. Oswaldo Cruz

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Section: Discussionmentioning

confidence: 99%

Trypanosoma cruzi: inhibition of alpha-hydroxyacid dehydrogenase isozyme II by N-allyl and N-propyl oxamates and their effects on intact epimastigotes

Chena¹,

Elizondo-Jiménez²,

Rodrı́guez-Páez³

et al. 2004

Mem. Inst. Oswaldo Cruz

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“…How could Tu target glucose metabolism? It has been well established that some adenosine analogs inhibit trypanosome glycolytic enzymes (18). The rationale behind this effort is that all but two of these enzymes contain adenosine-binding domains interacting with either NAD(H) or ADP/ATP.…”

Section: Discussionmentioning

confidence: 99%

The Adenosine Analog Tubercidin Inhibits Glycolysis in Trypanosoma brucei as Revealed by an RNA Interference Library

Drew

Morris

Wang

et al. 2003

Journal of Biological Chemistry

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We used an RNA interference (RNAi) library in a forward genetic selection to study the mechanism of toxicity of tubercidin (7-deazaadenosine) to procyclic Trypanosoma brucei. Following transfection of cells with an RNAibased genomic library, we used 5 M tubercidin to select a drug-resistant cell line. Surprisingly, we found in these resistant cells that the hexose transporters had been silenced. We subsequently found that silencing of hexokinase, a glycolytic enzyme, also yielded tubercidinresistant parasites. These observations suggested that glycolysis could be a target of tubercidin action and that RNAi silencing of glycolytic enzymes was gradual enough to allow the parasites to adapt to alternative sources of energy. Indeed, adaptation of procyclic trypanosomes to a glucose-independent metabolism by reduction of glucose in the culture medium caused tubercidin resistance. High pressure liquid chromatography analysis of glycolytic intermediates from parasites treated with tubercidin showed a dose-dependent increase in concentration of 1,3-bisphosphoglycerate, a substrate of phosphoglycerate kinase. Furthermore, tubercidin triphosphate inhibited recombinant T. brucei phosphoglycerate kinase activity in vitro with an IC 50 of 7.5 M. We conclude that 5 M tubercidin kills trypanosomes by targeting glycolysis, especially by inhibition of phosphoglycerate kinase.

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“…GAPDH has been targeted from protozoan parasites (27)(28)(29)(30), as the bloodstream forms rely solely on glycolysis for energy production (31,32). A number of mammalian GAPDH structures have also been solved, including rabbit * This work was supported in part by the Biotechnology and Biological Sciences Research Council.…”

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confidence: 99%

Structure of Insoluble Rat Sperm Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) via Heterotetramer Formation with Escherichia coli GAPDH Reveals Target for Contraceptive Design

Frayne

Taylor

Cameron

et al. 2009

Journal of Biological Chemistry

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Sperm glyceraldehyde-3-phosphate dehydrogenase has been shown to be a successful target for a non-hormonal contraceptive approach, but the agents tested to date have had unacceptable side effects. Obtaining the structure of the sperm-specific isoform to allow rational inhibitor design has therefore been a goal for a number of years but has proved intractable because of the insoluble nature of both native and recombinant protein. We have obtained soluble recombinant sperm glyceraldehyde-3-phosphate dehydrogenase as a heterotetramer with the Escherichia coli glyceraldehyde-3-phosphate dehydrogenase in a ratio of 1:3 and have solved the structure of the heterotetramer which we believe represents a novel strategy for structure determination of an insoluble protein.A structure was also obtained where glyceraldehyde 3-phosphate binds in the P s pocket in the active site of the sperm enzyme subunit in the presence of NAD. Modeling and comparison of the structures of human somatic and sperm-specific glyceraldehyde-3-phosphate dehydrogenase revealed few differences at the active site and hence rebut the long presumed structural specificity of 3-chlorolactaldehyde for the sperm isoform. The contraceptive activity of ␣-chlorohydrin and its apparent specificity for the sperm isoform in vivo are likely to be due to differences in metabolism to 3-chlorolactaldehyde in spermatozoa and somatic cells. However, further detailed analysis of the sperm glyceraldehyde-3-phosphate dehydrogenase structure revealed sites in the enzyme that do show significant difference compared with published somatic glyceraldehyde-3-phosphate dehydrogenase structures that could be exploited by structure-based drug design to identify leads for novel male contraceptives.Glyceraldehyde-3-phosphate dehydrogenase-S (GAPDS 3 in rat; GAPDH2 in human) is the sperm-specific isoform of GAPDH (1-3) and the sole GAPDH enzyme in sperm. GAPDS is highly conserved between species showing 94% identity between rat and mouse and 87% identity between rat and human. Within a particular species, GAPDS also shows significant sequence identity to its GAPDH paralogue, 70, 70, and 68% for rat, mouse, and human, respectively. The most striking difference between GAPDS and GAPDH is the presence of an N-terminal polyproline region in GAPDS, which is 97 residues in rat (accession number AJ297631), 105 in mouse (3), and 72 in human (2). GAPDS is restricted to the principal piece of the sperm flagellum (1, 2, 4) where it is localized to the fibrous sheath (5), an association proposed to be mediated via the N-terminal polyproline extension.GAPDS first came to prominence as a contraceptive target during the 1970s (6 -8). Investigations showed that treatment of sperm with ␣-chlorohydrin or a number of related compounds could inhibit GAPDS activity (9 -11), sperm motility (9 -13), and the fertilization of oocytes in vitro (14). The metabolite of these compounds, 3-chlorolactaldehyde (15-17), selectively inhibited GAPDS, having no effect on the activity of somatic cell GAPDH (18,19), pro...

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Glycolysis as a target for the design of new anti-trypanosome drugs

Cited by 210 publications

References 100 publications

Trypanosoma cruzi: inhibition of alpha-hydroxyacid dehydrogenase isozyme II by N-allyl and N-propyl oxamates and their effects on intact epimastigotes

Trypanosoma cruzi: inhibition of alpha-hydroxyacid dehydrogenase isozyme II by N-allyl and N-propyl oxamates and their effects on intact epimastigotes

The Adenosine Analog Tubercidin Inhibits Glycolysis in Trypanosoma brucei as Revealed by an RNA Interference Library

Structure of Insoluble Rat Sperm Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) via Heterotetramer Formation with Escherichia coli GAPDH Reveals Target for Contraceptive Design

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