New aspects of the inhibition of soybean lipoxygenase by alpha-tocopherol. Evidence for the existence of a specific complex.
The formation of alpha-tocopherol--lipoxygenase complex was elucidated using immobilized affinity purified soybean lipoxygenase and [D-3H]alpha-tocopherol. The alpha-tocopherol--lipoxygenase complex did not dissociate on addition of linoleic acid. Iodoacetate modified immobilized lipoxygenase did not form the complex with alpha-tocopherol. Lipoxygenase attached to an aminoethyl linoleyl Sepharose column was eluted by alpha-tocopherol. DL-alpha-Tocopherol acetate at a concentration of 3 X 10(-3) M inhibited 80% of linoleate oxidation by soybean lipoxygenase. The lipoxygenase--alpha-tocopherol complex did not give the usual soybean lipoxygenase antigenic pattern in immunodiffusion. Digestion of the [3H]alpha-tocopherol--lipoxygenase complex with proteolytic enzymes showed that most of the radioactivity is incorporated into one peptide.
D001666:Binding Sites, Antibody; D004800:Enzymes, Immobilized / Q000378:metabolism; D005779:Immunodiffusion; D016859:Lipoxygenase Inhibitors; D011485:Protein Binding; D013025:Glycine max / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D014810:Vitamin E / Q000494:pharmacology*
D004800:Enzymes, Immobilized; D016859:Lipoxygenase Inhibitors; D014810:Vitamin E