Kinetic and biochemical characterization of Trypanosoma evansi nucleoside triphosphate diphosphohydrolase

Weiss, Paulo Henrique Exterchoter; Batista, Franciane; Wagner, Glauber; Magalhães, Maria de Lourdes Borba; Miletti, Luiz Cláudio

doi:10.1016/j.exppara.2015.03.009

Cited by 4 publications

(3 citation statements)

References 56 publications

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“…Several studies have been performed to develop glucose-tolerant β-glucosidases. , An alternative strategy developed earlier consisted of glucose conversion into glucose-6-phosphate (G6P) followed by the investigation of β-glucosidase inhibition by G6P . The authors demonstrated that G6P does not inhibit the enzyme, but instead, G6P binds to the enzyme and causes conformational changes that culminate with increased thermal resistance, improved kinetic parameters, and glucose tolerance behavior .…”

Section: Introductionmentioning

confidence: 99%

Insights into Glucose-6-phosphate Allosteric Activation of β-Glucosidase A

Gomes

Silva

Lakkaraju

et al. 2021

J. Chem. Inf. Model.

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Second-generation ethanol production involves the use of agricultural and forestry waste as feedstock, being an alternative to the first-generation technology as it relies on low-cost abundant residues and does not affect food agriculture. However, the success of second-generation biorefineries relies on energetically efficient processes and effective enzyme cocktails to convert cellulose into fermentable sugars. β-glucosidases catalyze the last step on the enzymatic hydrolysis of cellulose; however, they are often inhibited by glucose. Previous studies demonstrated that glucose-6-phosphate (G6P) is a positive allosteric modulator of Bacillus polymyxa β-glucosidase A, improving enzymatic efficiency, providing thermoresistance, and imparting glucose tolerance. However, the precise molecular details of G6P-β-glucosidase A interactions have not yet been described so far. We investigated the molecular details of G6P binding into B. polymyxa β-glucosidase A through in silico docking using the site identification by ligand competitive saturation technology followed by site-directed mutagenesis studies, from which an allosteric binding site for G6P was identified. In addition, a mechanistic shift toward the transglycosylation reaction as opposed to hydrolysis was observed in the presence of G6P, suggesting a new role of G6P allosteric modulation of the catalytic activity of β-glucosidase A.

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Section: Introductionmentioning

confidence: 99%

Insights into Glucose-6-phosphate Allosteric Activation of β-Glucosidase A

Gomes

Silva

Lakkaraju

et al. 2021

J. Chem. Inf. Model.

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“…The enzyme has been characterized in many of the protozoan parasites and shows the same characteristic properties. Although there have been many inhibitors reported for the ecto-ATPase class of enzymes (de Souza Leite et al 2007;Weiss et al 2015), but these inhibitors do not show any promising inhibition of survival rate of parasites in the host. In addition, the activity of diphosphohydrolases is not affected by other ATPase and phosphatase inhibitors (Pereira-Neves et al 2014).…”

Section: Ectonucleotide Diphosphohydrolasementioning

confidence: 99%

Dependence of Leishmania parasite on host derived ATP: an overview of extracellular nucleotide metabolism in parasite

Arora

Kumar

2018

J Parasit Dis

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The widespread role of ATP in humans has been characterized as a substrate for metabolism as well as other key processes occurring in the human system. Hence, the ATP pool may also be accessible to the invaders, including Leishmania donovani (a protozoan parasite), considering the fact that ATP is not scarce at the cellular location where the pathogen resides. The protozoan parasite survives in the human host by utilizing purines from its extracellular environment, due to its inability to synthesize purines de novo. The purines are accessible in the form of nucleoside triphosphates (NTPs), for example, adenosine (ADO) in the form of ATP molecules. These NTPs are processed by the ecto-nucleotidases in the transmembrane region of the parasite, which are then transported inside via nucleoside/ nucleobase transporters belonging to the ENT family of & Ambak Kumar Rai

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“…These enzymes occur in several organisms, including Toxoplasma gondii [3], T. cruzi [4], T. evansi [5], Trichomonas vaginalis [6], and Leishmania infantum [7]. NTPDase from parasites can hydrolyze extracellular nucleoside tri- and diphosphate, thereby affecting the host's immune response [8–10].…”

Section: Introductionmentioning

confidence: 99%

Immobilization of NTPDase-1 fromTrypanosoma cruziand Development of an Online Label-Free Assay

Lima

Oliveira

Mariotini-Moura

et al. 2016

Journal of Analytical Methods in Chemistry

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The use of IMERs (Immobilized Enzyme Reactors) as a stationary phase coupled to high performance chromatographic systems is an interesting approach in the screening of new ligands. In addition, IMERs offer many advantages over techniques that employ enzymes in solution. The enzyme nucleoside triphosphate diphosphohydrolase (NTPDase-1) from Trypanosoma cruzi acts as a pathogen infection facilitator, so it is a good target in the search for inhibitors. In this paper, immobilization of NTPDase-1 afforded ICERs (Immobilized Capillary Enzyme Reactors). A liquid chromatography method was developed and validated to monitor the ICER activity. The conditions for the application of these bioreactors were investigated, and excellent results were obtained. The enzyme was successfully immobilized, as attested by the catalytic activity detected in the TcNTPDase-1-ICER chromatographic system. Kinetic studies on the substrate ATP gave K M of 0.317 ± 0.044 mmol·L−1, which still presented high affinity compared to in solution. Besides that, the ICER was stable for 32 days, enough time to investigate samples of possible inhibitors, including especially the compound Suramin, that inhibited 51% the enzyme activity at 100 µmol·L−1, which is in accordance with the data for the enzyme in solution.

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Kinetic and biochemical characterization of Trypanosoma evansi nucleoside triphosphate diphosphohydrolase

Cited by 4 publications

References 56 publications

Insights into Glucose-6-phosphate Allosteric Activation of β-Glucosidase A

Insights into Glucose-6-phosphate Allosteric Activation of β-Glucosidase A

Dependence of Leishmania parasite on host derived ATP: an overview of extracellular nucleotide metabolism in parasite

Immobilization of NTPDase-1 fromTrypanosoma cruziand Development of an Online Label-Free Assay

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