Plant and fungal lipoxygenases

Abstract

Lipoxygenases (LOX) are ubiquitous in plants, and some are important for resistance to pathogens. The soybean LOX (SLOX) remain the prototypes for studying lipoxygenation. The 3-D structures of SLOX-1 and -3 reveal an N-terminal beta-barrel and a C-terminal catalytic domain. The beta-barrel with 30 kDa of the N-terminal peptide chain of SLOX-1 can be separated from the catalytic domain (60 kDa), which is a functional lipoxygenase designated "mini-LOX." The 3-D structures have made it possible to predict and interpret effects of point mutations on catalytic parameters and on the substrate and position specificity and to identify the catalytic base as Fe3+-OH-. The first catalytic step, hydrogen abstraction at C-11 of linoleic acid by Fe3+-OH-, is associated with a very large isotope effect (20-80), which can be explained by the quantum-mechanical concept of hydrogen tunneling "through a 2-D potential barrier." The prosthetic iron of SLOX-3 can be extracted and the apoenzyme can incorporate Fe3+ and regain LOX activity. Plant LOX can be elicited by fungal pathogens. One fungal LOX has been cloned and characterized. It is secreted by the take-all fungus, and the enzyme may contribute to its detrimental actions on wheat roots. The primary structure of Mn-LOX shows that it belongs to the LOX gene family, the metal ligands appear to be homologous, and electron paramagnetic resonance (EPR) spectra suggest redox cycling between Mn2+ and Mn3+. The hypothetical catalytic base, Mn3+-OH-, abstracts the pro-S hydrogen at C-11, and molecular oxygen is inserted in a suprafacial way at C-11 and -13 in a 1:3 ratio. Mn-LOX catalyses the conversion of the 11S-hydroperoxide to 13R-hydroperoxylinoleic acid. Mn-LOX is a novel tool for studying lipoxygenation and plant-fungal interactions.