Using evolutionary changes to achieve species-specific inhibition of enzyme action — studies with triosephosphate isomerase

Gómez‐Puyou, Armando; Saavedra, Emma; Becker, Ingeborg; Zubillaga, Rafael A.; Rojo-Domlnguez, Arturo; Pérez‐Montfort, Ruy

doi:10.1016/1074-5521(95)90091-8

Cited by 70 publications

(72 citation statements)

References 62 publications

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“…Enzymatic activity assays. Enzymatic activity was determined following the conversion of glyceraldehyde 3-phosphate into dihydroxyacetone phosphate [7]. The decrease in absorbance at 340 nm was followed in a multicell Cary spectrophotometer at 25 °C.…”

Section: Inhibition Of Tctimmentioning

confidence: 99%

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3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies

et al. 2015

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Abstract:The current pharmacological Chagas disease treatments, using Nifurtimox or Benznidazole, show limited therapeutic results and are associated with potential side effects, OPEN ACCESSMolecules 2015, 20 14596 like mutagenicity. Using random screening we have identified new chemotypes that were able to inhibit relevant targets of the Trypanosoma cruzi. We found 3H-[1,2]dithioles with the ability to inhibit Trypanosoma cruzi triosephosphate isomerase (TcTIM). Herein, we studied the structural modifications of this chemotype to analyze the influence of volume, lipophilicity and electronic properties in the anti-T. cruzi activity. Their selectivity to parasites vs. mammalian cells was also examined. To get insights into a possible mechanism of action, the inhibition of the enzymatic activity of TcTIM and cruzipain, using the isolated enzymes, and the inhibition of membrane sterol biosynthesis and excreted metabolites, using the whole parasite, were achieved. We found that this structural framework is interesting for the generation of innovative drugs for the treatment of Chagas disease.

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Section: Inhibition Of Tctimmentioning

confidence: 99%

“…TcTIM is considered a potential target for anti-trypanosomal drugs [7][8][9]. This enzyme is involved in the glycolysis pathway of the parasite.…”

Section: Introductionmentioning

confidence: 99%

3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies

et al. 2015

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“…This proved to be the case. Following the idea that from measurements of the effect of a given reagent in homologous enzymes, it is possible to gain insight or determine the residue that is modified [6], we found that among triosephosphate isomerases that lack Cysl4, only those that have a Cys in position 217 are inhibited by a thiosulfonate that produces phenyl disulfides. Thus, the results illustrate that nonconserved residues can be used to find or produce species-specific inhibitors of homologous enzymes that exhibit a high level of selectivity.…”

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confidence: 99%

“…In more general terms, we studied whether various non-conserved residues in triosephosphate isomerase could represent potential targets for selective inhibition of homologous enzymes. Our leads in these studies were that some specific and non-specific sulfhydryl reagents inhibit the activity of triosephosphate isomerase from various species to different extents and that in rabbit triosephos-phate isomerase which has five Cys but lacks Cysl4, MeS0,-SMe produces a small inhibiting effect [6]. We therefore studied whether a higher inhibition could be produced in rabbit triosephosphate isomerase by larger derivatizing groups.…”

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confidence: 99%

“…We have suggested [6] that it is feasible to achieve species-specific inhibition of homologous enzymes with a high level of selectivity by targeting on amino acid residues that have not been conserved in evolution. In fact, inhibition of triosephosphate isomerases that possess a non-conserved Cys residue in position 14 was achieved by a cysteinereactive agent that produces methyl disulfides [7], i.e.…”

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Species‐Specific Inhibition of Homologous Enzymes by Modification of Nonconserved Amino Acids Residues

Garza‐Ramos

Pérez‐Montfort

Rojo‐Domínguez

et al. 1996

European Journal of Biochemistry

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The possibility of using non-conserved amino acid residues to produce selective inhibition of homologous enzymes from different species has been further explored with triosephosphate isomerase. S-phenylp-toluenethiosulfonate (MePhS0,-SPh), which produces phenyl disulfides with accessible Cys residues, inhibits the activity of rabbit triosephosphate isomerase. The inhibition is due to derivatization of one of the five Cys residues of rabbit triosephosphate isomerase. The effect of MePhS0,-SPh on triosephosphate isomerase from Saccharomyces cerevisiae, Escherichia coli, chicken and Schizosaccharomyces pombe was also determined. MePhS0,-SPh did not affect the activity of triosephosphate isomerase from S. cerevisiae and E. coli but it inhibited triosephosphate isomerase from chicken and S. pornhe. From an analysis of the Cys content of the various triosephosphate isomerases, it was evident that amongst the ones studied only those that have a Cys in position 217 (or in an equivalent position) were sensitive to MePhSOz-SPh. Methyl metanethiosulfonate (MeS0,-SMe), which produces methyl disulfides, had no effect on triosephosphate isomerases that lack Cys217 (S. cerevisiae and E. coli). In triosephosphate isomerases that have Cys217, MeS0,-SMe inhibited by 40-50% the activity of that from S. pornbe, 20-25% that from rabbit but had no effect on the chicken enzyme. In the three latter triosephosphate isomerases, MeS0,-SMe protected against the strong inhibiting action of MePhS0,-SPh. The latter observations suggest that MeS0,-SMe and MePhS0,-SPh derivatize the same Cys and that significant inhibition of activity requires perturbation by the relatively large phenyl group. The intrinsic fluorescence of rabbit triosephosphate isomerase that had been derivatized to a phenyl disulfide was almost identical to that of the native enzyme. Thus, modification of Cys217 did not produce gross structural alterations, albeit it brought about important kinetic alterations, i.e. a nearly fivefold increase in the K,, for glyceraldehyde 3-phosphate and a 65% decrease in V,,,,,. The effect of derivatizating Cys217 differs markedly from that produced by derivatization of Cysl4 (another non-conserved cysteine). The differences may be explained from their position in the three-dimensional structure of the enzyme.

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Energy Metabolism as an Antimalarial Drug Target

Jortzik

Becker

2011

Apicomplexan Parasites

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Using evolutionary changes to achieve species-specific inhibition of enzyme action — studies with triosephosphate isomerase

Cited by 70 publications

References 62 publications

3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies

3-H-[1,2]Dithiole as a New Anti-Trypanosoma cruzi Chemotype: Biological and Mechanism of Action Studies

Species‐Specific Inhibition of Homologous Enzymes by Modification of Nonconserved Amino Acids Residues

Energy Metabolism as an Antimalarial Drug Target

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