Crystal Structure of a Lipoxygenase in Complex with Substrate THE ARACHIDONIC ACID-BINDING SITE OF 8R-LIPOXYGENASE

被引:62
作者
Neau, David B. [2 ]
Bender, Gunes [1 ]
Boeglin, William E. [3 ]
Bartlett, Sue G. [1 ]
Brash, Alan R. [3 ]
Newcomer, Marcia E. [1 ]
机构
[1] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA
[2] Cornell Univ, Argonne Natl Lab, Dept Chem & Chem Biol, Northeastern Collaborat Access Team, Argonne, IL 60439 USA
[3] Vanderbilt Univ, Dept Pharmacol, Sch Med, Nashville, TN 37232 USA
基金
美国国家卫生研究院;
关键词
Arachidonic Acid (AA) (ARA); Eicosanoid Biosynthesis; Lipid Signaling; Lipoxygenase Pathway; Protein Structure; X-ray Crystallography; ACTIVE-SITE; ARACHIDONIC-ACID; FATTY-ACIDS; INHIBITION; OXYGENATION; SPECIFICITY; MODEL; STEREOSPECIFICITY; 8R-LIPOXYGENASE; DETERMINANTS;
D O I
10.1074/jbc.M114.599662
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: Lipoxygenases (LOX) catalyze the oxygenation of polyunsaturated fatty acids but generate distinct products from a common substrate. Results: We report the first structure of a LOX-substrate complex. Conclusion: The structure provides a context for understanding product specificity in enzymes that metabolize arachidonic acid. Significance: With roles in the production of potent lipid mediators, LOX are targets for drug design. 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 angstrom 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.
引用
收藏
页码:31905 / 31913
页数:9
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