Purification, characterization and kinetic properties of the Taenia solium glutathione S -transferase isoform 26.5�kDa

Plancarte, A.; Rendón, Juan L.; Landa, Abraham

doi:10.1007/s00436-004-1103-y

Cited by 17 publications

(13 citation statements)

References 33 publications

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“…A similar trend was reported earlier with temperature optima of around 40°C (Gupta et al 2007;Plancarte et al 2004) and loss of activity at higher temperatures.…”

Section: Discussionsupporting

confidence: 74%

Biochemical studies on glutathione S-transferase from the bovine filarial worm Setaria digitata

et al. 2011

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Setaria digitata is a filarial worm of the cattle used as a model system for antifilarial drug screening, due to its similarity to the human filarial parasites Wuchereria bancrofti and Brugia malayi. Since filarial glutathione S-transferase (GST) is a good biochemical target for antifilarial drug development, a study has been undertaken for the biochemical characterization of GST from S. digitata. Cytosolic fraction was separated from the crude S.digitata worm homogenate by ultracentrifugation at 100,000 g and subjected to ammonium sulfate precipitation followed by affinity chromatography using GSH-agarose column. The kinetic parameters K (m) and V (max) values with respect to GSH were 0.45 mM and 0.105 μmol min(-1) mL(-1) respectively. With respect to 1-chloro-2,4-dinitrobenzene, the K (m) and V (max) values were 1.21 and 0.117 μmol min(-1) mL(-1) respectively. The effect of temperature and pH on GST enzyme activity was studied. The protein retained its enzyme activity between 0°C and 40°C, beyond which it showed a decreasing tendency, and at 80°C, the activity was lost completely. The enzyme activity was varying with change in pH, and the maximum GST activity was observed at pH 7.5. Gel filtration chromatographic studies indicated that the protein has a native molecular mass of about 54 kDa. The single band of GST subunit appeared in sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to have molecular mass of ∼27 kDa. This shows that cytosolic S. digitata GST protein is homodimeric in nature.

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“…A similar trend was reported earlier with temperature optima of around 40°C (Gupta et al 2007;Plancarte et al 2004) and loss of activity at higher temperatures.…”

Section: Discussionsupporting

confidence: 74%

Biochemical studies on glutathione S-transferase from the bovine filarial worm Setaria digitata

et al. 2011

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“…The TsMF apparent K m value for GSH (0.164 mM) was lower than that reported for the T. solium cytoplasmic GST (TsGST 26.5 ; Plancarte et al 2004). In accordance with the pH activity and stability data, TsMF had a lower capacity to maintain the proper conformation of the active site in comparison with the TsGST 26.5 .…”

Section: Discussioncontrasting

confidence: 63%

“…At least two different soluble GSTs have been obtained for T. solium as for other helminths (Brophy and Barret 1990;Vibanco-Perez et al 2002;Plancarte et al 2004). The current work highlights the importance of this detoxification system in T. solium by proving the existence of GST activity in the parasite microsomes and its property to Glutathione S-transferase activity were assayed as described in "Materials and methods."…”

Section: Discussionmentioning

confidence: 97%

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Characterization of Taenia solium cysticerci microsomal glutathione S-transferase activity

2007

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Glutathione S-transferase activity has been shown to be associated with the microsomal fraction of Taenia solium. Electron microscopy and subcellular enzyme markers indicate the purity of the microsomal fraction that contains the glutathione S-transferase activity. T. solium microsomes were solubilized under conditions used to solubilize integral microsomal proteins. This procedure proved necessary to obtain enzymatic activity. To characterize this parasite enzyme activity, several substrates and inhibitors were used. The optimum activity for microsomal glutathione S-transferase was found to be pH 6.6, with a specific enzyme activity of 0.9, 0.1, 0.067, 0.03, and 0.05 micromol min(-1) mg(-1) with the substrates 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene, 4-hydroxynonenal, 2,4-hexadienal, and trans-2-nonenal, respectively. No activity of glutathione peroxidase was observed. T. solium microsomes had an appKm (GSH)=0.161 microM, appKm (CDNB)=14.5 microM, and appVmax of 0.15 and 27.9 micromol min(-1) mg(-1) for GSH and CDNB, respectively. T. solium microsomes were inhibited by several glutathione S-transferase enzyme inhibitors, and it was possible to establish a simple inhibition system as well as corresponding Ki's for each inhibitor. These results indicate that the T. solium microsomal glutathione S-transferase is different from the parasite cytoplasmic enzymes that catalyze similar reactions.

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“…In addition, the activity ranges with substrates such as ethacrynic acid, bromosulfophtalein, some alkenals, cumene hydroperoxide, and its sensitivities to inhibitors such as cibacron blue, bromosulfophtalein and chlorotriphenyltin, confirmed its mu nature. enzyme shares characteristics of M and A classes [62]. This finding suggests that both GSTs have great significance for the parasite survival [61].…”

Section: Gsh + Rx Gsr+ Hx (1)mentioning

confidence: 99%

Taenia solium: Antioxidant Metabolism Enzymes as Targets for Cestocidal Drugs and Vaccines

Vaca-Paniagua¹,

Torres-Rivera²,

Parra‐Unda³

et al. 2008

CTMC

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This review focuses in the role that antioxidant enzymes play in protection and other important physiological functions such as signal transduction, cell differentiation, growth and apoptosis. Parasites use these enzymes to evade ROS produced by the host immune response and for development inside the host. In the cestoda Taenia solium, three antioxidant enzymes have been studied: a cystosolic Cu,Zn superoxide dismutase that is a target of cestocidal drugs (bencimidazoles); a 2-Cys peroxiredoxin that is a regulatory enzyme of H(2)O(2), molecule essential for several physiological functions; and two isoforms of glutathione transferases that are immunological targets, since they protect immunized mice against cysticercosis. Moreover, all these enzymes are present in all stages of the parasite. These findings suggest that antioxidant enzymes have an important role in T. solium physiology and infection, therefore they might represent the Achilles' heel of the parasite.

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Purification, characterization and kinetic properties of the Taenia solium glutathione S -transferase isoform 26.5�kDa

Cited by 17 publications

References 33 publications

Biochemical studies on glutathione S-transferase from the bovine filarial worm Setaria digitata

Biochemical studies on glutathione S-transferase from the bovine filarial worm Setaria digitata

Characterization of Taenia solium cysticerci microsomal glutathione S-transferase activity

Taenia solium: Antioxidant Metabolism Enzymes as Targets for Cestocidal Drugs and Vaccines

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