Purification and partial characterization of glutathione transferase from the teleost Monopterus albus

Comp Biochem Physiol C Toxicol Pharmacol. 2008 Jan;147(1):96-100. doi: 10.1016/j.cbpc.2007.08.004. Epub 2007 Aug 22.

Abstract

Glutathione transferases (GSTs) catalyze the transfer of glutathione to a variety of xenobiotic and toxic endogenous compounds. GSTs are phase II biotransformation enzymes and are proposed as biomarkers of environmental pollution. In this study, a cytosolic glutathione transferase (maGST) was purified from liver of the freshwater fish Monopterus albus by affinity chromatography. The maGST appeared to be a homodimer composed of two subunits each with a molecular weight of 26 kDa. This maGST showed high activity towards the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl). Kinetic analysis with CDNB as substrate revealed a K(m) of 0.28 mM and V(max) of 15.68 micromol/min per mg of protein. It had maximum activity in the pH range 7.0-7.5, a broad optimum T(m) range of 30 degrees C-55 degrees C, and a high thermal stability with 77% of its initial activity at 45 degrees C. This high thermal stability of maGST could be related to the physiological adaptation of M. albus to high temperatures in tropical and subtropical environments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / metabolism
  • Animals
  • Chromatography, Affinity
  • Dimerization
  • Dinitrochlorobenzene / metabolism
  • Enzyme Stability
  • Fish Proteins / chemistry
  • Fish Proteins / isolation & purification*
  • Fish Proteins / metabolism
  • Glutathione Transferase / chemistry
  • Glutathione Transferase / isolation & purification*
  • Glutathione Transferase / metabolism
  • Hydrogen-Ion Concentration
  • Kinetics
  • Liver / enzymology*
  • Molecular Weight
  • Protein Denaturation
  • Smegmamorpha / metabolism*
  • Substrate Specificity
  • Temperature

Substances

  • Dinitrochlorobenzene
  • Fish Proteins
  • Glutathione Transferase
  • 4-Chloro-7-nitrobenzofurazan