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Published on: August 17, 2009
Isolation, purification, and characterization of glutathione S-transferase from oat (Avena sativa) seedlings
V T Zachariah1, N Walsh-Sayles, B R Singh
1Department of Chemistry and Biochemistry, and Center for Marine Science and Technology, University of Massachusetts Dartmouth, 02747, USA.
Abstract:
Glutathione S-transferase (GST) from oat seedlings was purified by ammonium sulfate precipitation and glutathione (GSH) affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of two major protein subunits with molecular masses of 29 and 31 kDa, respectively. Isoelectric focusing revealed a major band with pI of 3.43 and a minor band with pI of 7.42. Kinetic analysis with respect to 1-chloro-2,4-dinitrobenzene (CDNB) as substrate revealed a Km of 1.18 mM and Vmax of 0.94 micromol/min and a specific activity of 17.96 micromol/min/mg. Inhibition studies indicated that oat GST is strongly inhibited by chlorophyllin, hemin, and anthocyanin and only weakly by bilirubin and biliverdin.
