Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 877, Issue 26, Pages 2863-2872Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2009.03.007
Keywords
Oxidative lipidomics; Cardiolipin; Phosphatidylserine; Phospholipid hydroperoxides; Mass-spectrometry; Cytochrome c; Apoptosis
Funding
- NIH [HL70755]
- NIOSH [OH008282, HD057587, NS061817]
- DAMD [17-01-2-637]
- Pennsylvania Department of Health [SAP 4100027294, AHA-0535365N]
- Human Frontier Science Program [NORA 927Z1LU, NORA NTRC927ZGFY]
- NANOMMUNE [214281 FP7-NMP-2007-SMALL-1]
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Oxidation of two anionic phospholipids - cardiolipin (CL) in mitochondria and phosphatidylserine (PS) in extramitochondrial compartments - is important signaling event, particularly during the execution of programmed cell death and clearance of apoptotic cells. Quantitative analysis of CL and PS oxidation products is central to understanding their molecular mechanisms of action. We combined the identification of diverse phospholipid molecular species by ESI-MS with quantitative assessments of lipid hydroperoxides using a fluorescence HPLC-based protocol. We characterized CL and PS oxidation products formed in a model system (cyt c/H(2)O(2)), in apoptotic cells (neurons, pulmonary artery endothelial cells) and mouse lung under inflammatory/oxidative stress conditions (hyperoxia, inhalation of single walled carbon nanotubes). Our results demonstrate the usefulness of this approach for quantitative assessments, identification of individual molecular species and structural characterization of anionic phospholipids that are involved in oxidative modification in cells and tissues. (C) 2009 Elsevier B.V. All rights reserved.
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