Oxidative Lipidomics of Apoptosis: Quantitative Assessment of Phospholipid Hydroperoxides in Cells and Tissues

Details

• Personal Author:
  Amoscato AA ; Bayir H ; Dekosky ST ; Greenberger JS ; Hamilton R ; Kagan VE ; Kochanek PM ; Lysytsya A ; Ritov VB ; Tyurin VA ; Tyurina YY
• Description:
  Oxidized phospholipids play essential roles in execution of mitochondrial stage of apoptosis and clearance of apoptotic cells. The identification and quantification of oxidized phospholipids generated during apoptosis can be successfully achieved by oxidative lipidomics. With this approach, diverse molecular species of phospholipids and their hydroperoxides are identified and characterized by soft-ionization mass-spectrometry techniques such as electrospray ionization (ESI). Quantitative assessment of lipid hydroperoxides is performed by fluorescence HPLC-based protocol. The protocol is based on separation of phospholipids using two-dimensional-high-performance thin-layer chromatography (2-D-HPTLC). Phospholipids are hydrolyzed using phospholipase A2. The fatty acid hydroperoxides (FA-OOH) released is quantified by a fluorometric assay using Amplex red reagent and microperoxidase-11 (MP-11). Detection limit of this protocol is 1-2 pmol of lipid hydroperoxides. Lipid arrays vs. oxidized lipid arrays can be performed by comparing the abundance of phospholipids with the abundance of oxidized phospholipids. Using oxidative lipidomics approach we show that the pattern of phospholipid oxidation during apoptosis is nonrandom and does not follow their abundance in several types of cells undergoing apoptosis and a variety of disease states. This has important implications for evaluation of apoptosis in vivo. The anionic phospholipids, cardiolipin (CL) and phosphatidylserine (PS), are the preferred peroxidation substrates. [Description provided by NIOSH]
• Subjects:
  Chromatography Lipids Occupational Health Oxidants Safety
• Keywords:
  Alzheimer's Disease Apoptosis Author Keywords: Phospholipid Hydroperoxides Cardiolipin Controlled Cortical Impact Cytochrome C ESI-MS Fluorescence HPLC Gamma-irradiation Lipid Peroxidation Oxidation Oxidative Processes Phosphatidylserine Phospholipids

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• ISBN:
  9781588297105
• ISSN:
  1064-3745
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; Pennsylvania
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  353-374
• Volume:
  610
• NIOSHTIC Number:
  nn:20057827
• Citation:
  Methods Mol Biol 2010 Jan; 610:353-374
• Editor(s):
  Uppu RM; Murthy SN; Pryor WA; Parinandi NL
• Federal Fiscal Year:
  2010
• Performing Organization:
  University of Pittsburgh at Pittsburgh
• Peer Reviewed:
  False
• Start Date:
  20050701
• Source Full Name:
  Methods in Molecular Biology. Free Radicals and Antioxidant Protocols
• End Date:
  20160630
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:3ce6d8161507d0a2a72c60f56ee135298971146157665fef52bc00451c6324b4dee7c7007a3cbb590b2b1bd894245f83c1ed8b4451c6c22251579ebb5580aa52
• Download URL:
  https://stacks.cdc.gov/view/cdc/215044/cdc_215044_DS1.pdf
• File Type:
  [PDF - 460.71 KB ]