Development of MALDI-TOF methodology for selective MS-analysis and imaging of cardiolipins in lipid extracts and tissues

Details

• Personal Author:
  Amoscato A ; Bayir H ; Kagan VE ; Pitt BR ; Sparvero LJ
• Description:
  Cardiolipin (CL) is an anionic phospholipid that plays a key role in mitochondrial electron transport but is also highly susceptible to oxidative stress. The molecular diversity of endogenous cardiolipins in some tissues and the low abundance of its individual species, particularly oxidized derivatives, makes in situ analysis and imaging by MALDI-Mass Spectrometry (MALDI-MS) challenging. The goal of this study was to develop a method to selectively analyze CL using existing CL probes in a novel way, to decrease CL mass spectral complexity. Nonyl Acridine Orange (NAO) is a commonly used fluorescent stain for cardiolipin. We have used NAO as a novel MALDI matrix, and have found that it allowed detection of anionic phospholipids, including cardiolipin in the negative ion mode. With careful laser tuning, NAO induced prompt decay of cardiolipin selectively - but not other phospholipids - at very high efficiencies. From these selective prompt decay products, much information about the structure of cardiolipin can be determined. Pseudo- MS3 is possible on a MALDI-TOF instrument by doing MS/MS on these products of cardiolipin. We have also extended these findings to include oxidized species of CL's and have determined that NAO also induced prompt decay of oxidized CL. Prompt fragments are seldom seen with dihydroxy-benzoic acid (DHB) which has been employed as a standard matrix for lipid analysis in the negative ion mode. Our results indicate that NAO can be utilized as a discerning matrix for the assessment of CL in lipid extracts from cells and the distribution of CL - particularly in tissues with a high diversity of CL molecular species and their oxidation products - by taking full advantage of the unique ability to induce prompt decay of this phospholipid with NAO. [Description provided by NIOSH]
• Subjects:
  Diagnosis Lipids Mass Spectrometry Occupational Health Respiratory System Risk Assessment Risk Factors Safety Toxicology
• Keywords:
  Biological-effects Cell-biology Cell-function Cell-morphology Cell-transformation Cellular-reactions Diagnostic-techniques Inhalation-studies Mass-spectrometry Medical-research Oxidation Oxidative-processes Phospholipids Physical-properties Physiological-effects Risk-analysis Risk-factors Statistical-analysis Tissue-culture

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• ISSN:
  1096-6080
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Abstract or Summary
• Place as Subject:
  District of Columbia ; OSHA Region 3 ; Pennsylvania ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  120
• NIOSHTIC Number:
  nn:20038522
• Citation:
  Toxicologist 2011 Mar; 120(Suppl 2):275
• Federal Fiscal Year:
  2011
• NORA Priority Area:
  Manufacturing
• Performing Organization:
  University of Pittsburgh at Pittsburgh
• Peer Reviewed:
  False
• Start Date:
  20050701
• Source Full Name:
  The Toxicologist. Society of Toxicology 50th Annual Meeting and ToxExpo, March 6-10, 2011, Washington, DC
• Supplement:
  2
• End Date:
  20160630
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:cdc305863af33c433974eac74d00876dabb1d80fdbb0306c358997b47e69e2927f7039071950d1d6331340f3a8c8e0dc64d30ff8bc696f31ae5c2d195beb25e5
• Download URL:
  https://stacks.cdc.gov/view/cdc/192207/cdc_192207_DS1.pdf
• File Type:
  [PDF - 1.09 MB ]