caa a22 4500 445339519 CHVBK 20180317142807.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s00436-010-2227-x doi (NATIONALLICENCE)springer-10.1007/s00436-010-2227-x Biochemical studies on glutathione S-transferase from the bovine filarial worm Setaria digitata [Elektronische Daten] [Lakshmy Srinivasan, Nisha Mathew, Twinkle Karunan, Kalyanasundaram Muthuswamy] 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 Km and Vmax values with respect to GSH were 0.45mM and 0.105μmolmin−1mL-1 respectively. With respect to 1-chloro-2,4-dinitrobenzene, the Km and Vmax values were 1.21 and 0.117μmolmin−1mL−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 pH7.5. Gel filtration chromatographic studies indicated that the protein has a native molecular mass of about 54kDa. The single band of GST subunit appeared in sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to have molecular mass of ∼27kDa. This shows that cytosolic S. digitata GST protein is homodimeric in nature. Springer-Verlag, 2010 Srinivasan Lakshmy Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut Mathew Nisha Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut Karunan Twinkle Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut Muthuswamy Kalyanasundaram Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut Parasitology Research Springer-Verlag 109/1(2011-07-01), 213-219 0932-0113 109:1<213 2011 109 436 https://doi.org/10.1007/s00436-010-2227-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-010-2227-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Srinivasan Lakshmy Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut NATIONALLICENCE 700 1- Mathew Nisha Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut NATIONALLICENCE 700 1- Karunan Twinkle Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut NATIONALLICENCE 700 1- Muthuswamy Kalyanasundaram Vector Control Research Centre (Indian Council of Medical Research), 605006, Indira Nagar, Pondicherry, India aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/1(2011-07-01), 213-219 0932-0113 109:1<213 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer