Immunological comparison of lipoxygenase isozymes-1 and -2 with soybean seedling lipoxygenases

Abstract

An affinity-purified polyclonal antibody against soybean seed lipoxygenase-2 was prepared and used to characterize the immunological relatedness of lipoxygenase isozymes 1 and 2 and lipoxygenases from soybean seedling roots, hypocotyls, leaves, and cotyledons. All soybean lipoxygenases tested cross-reacted with the anti-lipoxygenase-2. Cross-reactivity of seed-derived lipoxygenases was evidenced by formation of a line of identity in double-diffusion tests, by positive results on an immunoblot, and by antibody precipitation of enzyme activity. Levels of anti-lipoxygenase-2, which inhibited lipoxygenase-2 activity, had no effect on lipoxygenase-1 activity. Root, hypocotyl, and leaf lipoxygenases did not form precipitation lines in double-diffusion tests but the anti-lipoxygenase-2 did inhibit and precipitate lipoxygenase activity from these sources as well as cross-react on immunoblots. All the cross-reactive lipoxygenases examined were found to have the same apparent molecular weight. Lipoxygenase activity found in soybean seedling roots, hypocotyls, leaves, and cotyledons is associated with proteins which are all immunologically related to the seed-derived enzymes.