Structural Flexibility of the N-terminal β-Barrel Domain of 15-Lipoxygenase-1 Probed by Small Angle X-ray Scattering. Functional Consequences for Activity Regulation and Membrane Binding

Abstract

Mammalian lipoxygenases form a heterogeneous family of lipid peroxidizing enzymes, which have been implicated in synthesis of inflammatory mediators, in cell development and in the pathogenesis of various diseases (atherosclerosis, osteoporosis) with major health political importance. The crystal structures of two plant lipoxygenase isoforms have been solved and X-ray coordinates for an inhibitor complex of the rabbit 15-lipoxygenase-1 are also accessible. Here, we investigated the solution structure of the ligand-free rabbit 15-lipoxygenase-1 by small angle X-ray scattering. From the scattering profiles we modeled the solution structure of the enzyme using two independent ab initio approaches. Preliminary experiments indicated that at low protein concentrations (<1mg/ml) and at 10 degrees C the enzyme is present as hydrated monomer. Superposition of the high resolution crystal structure and our low resolution model of the solution structure revealed two major differences. (i) Although the two models are almost perfectly superimposed in the region of the catalytic domain the solution structure is stretched out in the region of the N-terminal beta-barrel domain and exhibits a bigger molecular volume. (ii) There is a central bending of the enzyme molecule in the solution structure, which does not show up in the crystal structure. Both structural peculiarities may be explained by a high degree of motional freedom of the N-terminal beta-barrel domain in aqueous solutions. This interdomain movement may be of functional importance for regulation of the catalytic activity and membrane binding.