title: Molecular speciation and dynamics of oxidized triacylglycerols in lipid droplets: Mass spectrometry and coarse-grained simulations.
abstract: Lipid droplets (LDs) are ubiquitous and physiologically active organelles regulating storage and mobilization of lipids in response to metabolic demands. Among the constituent LD neutral lipids, such as triacylglycerols, cholesterol esters, and free fatty acids, oxidizable polyunsaturated molecular species may be quite abundant, yet the structural and functional roles of their oxidation products have not been studied. Our previous work documented the presence of these peroxidized species in LDs. Assuming that hydrophilic oxygen-containing functionalities may markedly change the hydrophobic/hydrophilic molecular balance, here we utilized computational modeling to test the hypothesis that lipid peroxidation causes redistribution of lipids between the highly hydrophobic core and the polar surface (phospho)lipid monolayer-the area enriched with integrated enzymatic machinery. Using quantitative liquid chromatography/mass spectrometry, we characterized molecular speciation of oxTAGs in LDs of dendritic cells in cancer and hypoxic trophoblasts cells as two cellular models associated with dyslipidemia. Among the many types of oxidized lipids identified, we found that oxidatively truncated forms and hydroxyl derivatives of TAGs were the prevailing oxidized lipid species in LDs in both cell types. Using coarse-grained molecular dynamics (CG-MD) simulations we established that lipid oxidation changed their partitioning whereby oxidized lipids migrated into the outer monolayer of the LD, where they can affect essential metabolic pathways and undergo conversions, possibly leading to the formation of oxygenated lipid mediators.
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15219, USA; Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA 15219, USA; Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Dariush
authors.initials: D
authors.lastname: Mohammadyani
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15219, USA; Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Tyurin
authors.affiliation: Magee-Womens Research Institute, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: O'Brien
authors.affiliation: Magee-Womens Research Institute, University of Pittsburgh, Pittsburgh, PA 15219, USA.
authors.forename: Yoel
authors.initials: Y
authors.lastname: Sadovsky
authors.affiliation: The Wistar Institute, Philadelphia, PA 19104, USA.
authors.forename: Dmitry I
authors.initials: DI
authors.lastname: Gabrilovich
authors.affiliation: Division of Metabolic and Vascular Health, Medical School, University of Warwick, Coventry CV4 7AL, UK.
authors.forename: Judith
authors.initials: J
authors.lastname: Klein-Seetharaman
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15219, USA; Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15219, USA. Electronic address: kagan@pitt.edu.
authors.forename: Valerian E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D002853:Chromatography, Liquid; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D066292:Lipid Droplets / Q000737:chemistry*; D008055:Lipids / Q000032:analysis; D008223:Lymphoma / Q000378:metabolism* / Q000473:pathology; D056004:Molecular Dynamics Simulation; D010084:Oxidation-Reduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014280:Triglycerides / Q000737:chemistry*; D014327:Trophoblasts; D014407:Tumor Cells, Cultured
chemical_list: D008055:Lipids; D014280:Triglycerides
keywords: Cancer; Hypoxia; Lipid droplet; Lipid oxidation; Neutral lipids
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