Elucidation of allosteric behavior and enzyme-effector complexes of human 15-lipoxygenase-2 through hydrogen-detuerium exchange mass spectrometry

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Amy Musgrave (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: The enzymatic activity of human 15-lipoxygenase-2 (15-LOX-2) has been implicated in human inflammatory diseases such as atherosclerotic cardiovascular disease and plays an important role in maintaining homeostasis. 15-LOX-2 is one of six human lipoxygenase enzyme that catalyzes the (per)oxidation of fatty acids to produce both pro- and anti-inflammatory cellular signaling molecules.1 Despite its importance , current anti-inflammatories available on the market do not target lipoxygenase derived inflammation. There are three types of regulation of 15-LOX-2 activity: small allosteric regulators , selective inhibitors , and protein-protein/protein-membrane interactions. The first of these types of regulation is the focus of this study. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) allows for structural analysis of the complex formed with small allosteric regulators and selective inhibitors of 15-LOX-2. Using HDX-MS , we have shown excellent coverage of the primary sequence of 15-LOX-2 with dynamic properties comparable to previous reports. Further , in the presence of oleyl sulfate , a small molecule effector , regions of the protein were resolved to show alterations in protein flexibility. This data provides a basis for 15-LOX-2 regulation.

Additional Information

Publication

Thesis

Language: English

Date: 2019

Keywords

lipoxygenase, allostery, hydrogen-deuterium exchange (HDX), HDX-MS, 15-LOX-2

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