Crystal structure of a lipoxygenase in complex with substrate: the arachidonic acid-binding site of 8R-lipoxygenase.
Publication Type:Journal Article
Source:J Biol Chem, Volume 289, Issue 46, p.31905-13 (2014)
Keywords:Animals, Arachidonate Lipoxygenases, Arachidonic Acid, Binding Sites, Catalysis, Crystallography, X-Ray, Humans, Inflammation, Iron, Lipids, Models, Molecular, Mutagenesis, Mutation, Oxygen, Protein Binding, Protein Conformation, Rabbits, Swine
<p>Lipoxygenases (LOX) play critical roles in mammalian biology in the generation of potent lipid mediators of the inflammatory response; consequently, they are targets for the development of isoform-specific inhibitors. The regio- and stereo-specificity of the oxygenation of polyunsaturated fatty acids by the enzymes is understood in terms of the chemistry, but structural observation of the enzyme-substrate interactions is lacking. Although several LOX crystal structures are available, heretofore the rapid oxygenation of bound substrate has precluded capture of the enzyme-substrate complex, leaving a gap between chemical and structural insights. In this report, we describe the 2.0 Å resolution structure of 8R-LOX in complex with arachidonic acid obtained under anaerobic conditions. Subtle rearrangements, primarily in the side chains of three amino acids, allow binding of arachidonic acid in a catalytically competent conformation. Accompanying experimental work supports a model in which both substrate tethering and cavity depth contribute to positioning the appropriate carbon at the catalytic machinery. </p>