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

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Abstract

Glutathione S-transferases are major phase II detoxification enzymes. Taenia solium, a parasite of humans and pigs, is exposed to toxic products. The aim of this work was to purify and characterize a T. solium glutathione S-transferase isoform of 26.5 kDa (SGST26.5) in order to obtain its kinetic parameters. Homogeneous SGST26.5 was obtained by a simple purification procedure. SGST26.5 showed a pI of 7.07, and a native M r of 60 kDa with 26.5 kDa subunits. The optimum activity for SGST26.5 was found at pH 6.5–7.0 in the range 10–42°C. SGST26.5 had a specific enzyme activity of 78, 7.1, 6.6, and 0.7 μM min−1 mg−1 with CDNB, 1,2-dichloro-4-nitrobenzene, 2,4-hexadienal and trans-2-nonenal as substrates, respectively. It also had a k cat/K m(CDNB)=2.15×103 M−1 s−1, k cat/K m(GSH)=4.5×103 M−1 s−1 and V max for GSH and CDNB=74 and 77 μM min−1 mg−1, respectively. SGST26.5 was inhibited in a noncompetitive form by cibacron blue, bromosulfophthalein and triphenyltin chloride. Inhibition studies as a function of inhibitor concentration show that the enzyme is a homodimer. Bireactant system analysis show that it follows an ordered sequential mechanism.

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Acknowledgements

This work was financed by the Consejo Nacional de Ciencia y Tecnología, México (Conacyt 27556-M) and Dirección General de Asuntos del Personal Académico, México) DGAPA IN212600. The authors wish to thank Dr. Gerardo Medina for valuable English corrections.

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Authors and Affiliations

  1. Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, UNAM, Circuiro interior, 04510, Mexico City, Mexico

    A. Plancarte & A. Landa

  2. Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, UNAM. Circuiro interior, 04510, Mexico City, Mexico

    J. L. Rendon

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  1. A. Plancarte
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Correspondence to A. Plancarte.

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Plancarte, A., Rendon, J.L. & Landa, A. Purification, characterization and kinetic properties of the Taenia solium glutathione S-transferase isoform 26.5 kDa. Parasitol Res 93, 137–144 (2004). https://doi.org/10.1007/s00436-004-1103-y

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