Abstract |
The enzyme encoded by the ALOX15B gene has been linked to the development of atherosclerotic plaques in humans and in a mouse model of hypercholesterolemia. In vitro, these enzymes, which share 78% sequence identity, generate distinct products from their substrate arachidonic acid: the human enzyme, a 15-S-hydroperoxy product; and the murine enzyme, an 8-S-product. We probed the activities of these enzymes with nanodiscs as membrane mimics to determine whether they can access substrate esterified in a bilayer and characterized their activities at the membrane interface. We observed that both enzymes transform phospholipid-esterified arachidonic acid to a 15-S-product. Moreover, when expressed in transfected HEK cells, both enzymes result in significant increases in the amounts of 15-hydroxyderivatives of eicosanoids detected. In addition, we show that 15-LOX-2 is distributed at the plasma membrane when the HEK293 cells are stimulated by the addition Ca(2+) ionophore and that cellular localization is dependent upon the presence of a putative membrane insertion loop. We also report that sequence differences between the human and mouse enzymes in this loop appear to confer distinct mechanisms of enzyme-membrane interaction for the homologues.
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Authors | Gunes Bender, Erin E Schexnaydre, Robert C Murphy, Charis Uhlson, Marcia E Newcomer |
Journal | The Journal of biological chemistry
(J Biol Chem)
Vol. 291
Issue 37
Pg. 19413-24
(09 09 2016)
ISSN: 1083-351X [Electronic] United States |
PMID | 27435673
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Copyright | © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. |
Chemical References |
- Arachidonic Acids
- Calcium Ionophores
- ALOX15B protein, human
- Arachidonate 15-Lipoxygenase
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Topics |
- Animals
- Arachidonate 15-Lipoxygenase
(chemistry, genetics, metabolism)
- Arachidonic Acids
(chemistry, genetics, metabolism)
- Atherosclerosis
(enzymology, genetics)
- Calcium Ionophores
(pharmacology)
- Cell Membrane
(chemistry, enzymology, genetics)
- Disease Models, Animal
- HEK293 Cells
- Humans
- Mice
- Protein Structure, Secondary
- Protein Transport
(drug effects)
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