Isolation, purification, and characterization of glutathione S-transferase from oat (Avena sativa) seedlings.
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.
Zachariah
V T
VT
Department of Chemistry and Biochemistry, and Center for Marine Science and Technology, University of Massachusetts Dartmouth, 02747, USA.
Walsh-Sayles
N
N
Singh
B R
BR
D000872:Anthocyanins / Q000494:pharmacology; D018554:Avena / Q000737:chemistry / Q000201:enzymology*; D001667:Binding, Competitive; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D010944:Plants / Q000737:chemistry / Q000201:enzymology; D011758:Pyrroles / Q000494:pharmacology; D045725:Tetrapyrroles
D000872:Anthocyanins; D011758:Pyrroles; D045725:Tetrapyrroles; D005982:Glutathione Transferase