Understanding the molecular mechanism of the Ala-versus-Gly concept controlling the pProduct specificity in reactions catalyzed by lipoxygenases

Lipoxygenases (LOXs) are a family of enzymes that catalyze the highly specific hydroperoxidation of polyunsaturated fatty acids, such as arachidonic acid. Different stereo- or/and regioisomer hydroperoxidation products lead later to different metabolites that exert opposite physiological effects in...

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Bibliographic Details
Authors: Saura Martínez, Patricia, Suardiaz Del Rio, Reynier|||0000-0002-1035-9020, Masgrau, Laura|||0000-0003-4495-508X, González-Lafont, Àngels|||0000-0003-0729-2483, Rosta, Edina, Lluch, José M.|||0000-0002-7536-1869
Format: article
Publication Date:2017
Country:España
Institution:Universitat Autònoma de Barcelona
Repository:Dipòsit Digital de Documents de la UAB
Language:English
OAI Identifier:oai:ddd.uab.cat:225211
Online Access:https://ddd.uab.cat/record/225211
https://dx.doi.org/urn:doi:10.1021/acscatal.7b00842
Access Level:Open access
Keyword:Coral 8R-lipoxygenase
Ala-versus-Gly concept
Enzyme catalysis
Arachidonic acid
QM/MM calculations
Molecular dynamics simulations
Free energy calculations
Description
Summary:Lipoxygenases (LOXs) are a family of enzymes that catalyze the highly specific hydroperoxidation of polyunsaturated fatty acids, such as arachidonic acid. Different stereo- or/and regioisomer hydroperoxidation products lead later to different metabolites that exert opposite physiological effects in the animal body and play a central role in inflammatory processes. The Gly-Ala switch of a single residue is crucial for the stereo- and regiocontrol in many lipoxygenases. Herein, we have combined molecular dynamics simulations with quantum mechanics/molecular mechanics calculations to study the hydrogen abstraction step and the molecular oxygen addition step of the hydroperoxidation reaction of arachidonic acid catalyzed by both wild-type Coral 8R-LOX and its Gly427Ala mutant. We have obtained a detailed molecular understanding of this Ala-versus-Gly concept. In wild type, molecular oxygen adds to C of arachidonic acid with an R stereochemistry. In the mutant, Ala427 pushes Leu385, blocks the region over C, and opens an oxygen access channel now directed to C, where molecular oxygen is added with an S stereochemistry. Thus, the specificity turns out to be dramatically inverted. Since Leu385 is highly conserved among many lipoxygenase isoforms, this mechanism can be general, and we propose that the presence of such type of bulky and hydrophobic residues can be key in controlling the extreme regio- and stereospecificity of lipoxygenases and, as a consequence, their physiological effects.