Demonstration by EPR spectroscopy of the functional role of iron in soybean lipoxygenase-1.
1. The EPR spectrum at 15 degrees K of soybean lipoxygenase-1 in borate buffer pH 9.0 has been studied in relation to the presence of substrate (linoleic acid), product (13-L-hydroperoxylinoleic acid) and oxygen. 2. The addition of 13-L-hydroperoxylinoleic acid to lipoxygenase-1 at pH 9.0 gives rise to the appearance of EPR lines at g equals 7.5, 6.2, 5.9 and 2.0, and an increased signal at g equals 4.3. 3. In view of the effect of the end product on both the kinetic lag period of the aerobic reaction and the fluorescence of the enzyme, it is concluded that 13-L-hydroperoxylinoleic acid is required for the activation of soybean lipoxygenase-1. Thus it is proposed that the enzyme with iron in the ferric state is the active species. 4. A reaction scheme is presented in which the enzyme alternatingly exists in the ferric and ferrous states for both the aerobic and anaerobic reaction.
D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation; D008041:Linoleic Acids; D008084:Lipoxygenase / Q000378:metabolism*; D008956:Models, Chemical; D010100:Oxygen; D010545:Peroxides / Q000494:pharmacology; D010944:Plants / Q000201:enzymology*; D011485:Protein Binding; D013025:Glycine max
D008041:Linoleic Acids; D010545:Peroxides; D008084:Lipoxygenase; D010100:Oxygen