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Lipid Mass Spectrometry in the Cardiovascular System Plasmalogen Targeting

Lipid Mass Spectrometry in the Cardiovascular System Plasmalogen Targeting by Reactive Chlorinating Species Derived from Phagocytes in Infarcted Myocardium David Ford, Ph.D. Department of Biochemistry and Molecular Biology St. Louis University School of Medicine.

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Lipid Mass Spectrometry in the Cardiovascular System Plasmalogen Targeting

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  1. Lipid Mass Spectrometry in the Cardiovascular System Plasmalogen Targeting by Reactive Chlorinating Species Derived from Phagocytes in Infarcted Myocardium David Ford, Ph.D. Department of Biochemistry and Molecular Biology St. Louis University School of Medicine

  2. MOLECULAR SUBCLASSES OF ETHANOLAMINE GLYCEROPHOSPHOLIPIDS O H H O C C H O CH C H O C C C H 15 31 2 15 31 14 29 O O O O C C H O C C H O C C H 19 31 19 31 19 31 O O O + + + O P O CH CH NH O P O CH CH NH O P O CH CH NH 3 2 2 - 3 3 2 2 2 2 - - O O O ALKYL ETHER DIACYL PLASMALOGEN (Plasmanylethanolamine) (Phosphatidylethanolamine) (Plasmenylethanolamine)

  3. Plasmalogen Targeting by Reactive Chlorinating Species Derived from Phagocytes Cl Schiff base adducts O C C C H 14 29 H H 2-Chlorohexadecanal ( -Chloro-fatty aldehyde) a + H H H O C C C 29 14 +H O , Cl O 2 2 OH O C R 1 Myeloperoxidase O CH O LPC chlorohydrins 3 O C R 1 CH O P O (CH ) N 3 2 2 + CH O 3 CH O 3 Plasmenylcholine CH O P O (CH ) N 3 2 2 (Plasmalogen) + CH O 3 Lysophosphatidylcholine

  4. GC-MS analysis by negative ion electron capture of the PFB oxime of 2-chlorohexadecanal produced from RCS attack of plasmalogens in infarcted myocardium

  5. 2-Chlorohexadecanal in infarcted myocardium D 1 day LAD Occlusion Mid-Ventricle 1 day sham surgery E Mid-Ventricle

  6. Concomitant accumulation of neutrophils and 2-chlorohexadecanal

  7. Schiff Base Formation O O O R R 2 1 O O O O O O P O - R R O 2 N H 1 2 O O Phosphatidylethanolamine (PE) O P O + - O N H2O O Cl C H Cl C H 14 29 14 29 N-2-ClHDA-PE 2-Chlorohexadecanal (2-ClHDA)

  8. 2-ClHDA Forms Schiff Bases with di16:0-PE

  9. O O O C C H 1 5 3 1 C H C O O 1 5 3 1 O P O - [ M - H ] = m / z 9 1 2 N H O C H 1 4 2 9 m / z 3 6 2 N-2-ClHDA-PE Schiff Base Adducts in HCAEC Incubate HCAEC with 25 mM 2-ClHDA Stabilize Schiff bases with NaCNBH3 Detect and quantify Schiff base adducts by LC-MS/MS (SRM) SRM = [M-H]- m/z 362

  10. N-2-ClHDA-PE Schiff Base Adducts in Human Coronary Artery Endothelial Cells

  11. Secondary RCS Attack of Plasmalogens: Production of LPC Chlorohydrins

  12. CID of m/z 596 (18 carbon LPC Chlorohydrin)

  13. Detection of LPC chlorohydrins in HCAEC treated with HOCl: Neutral Loss Scanning of m/z 95 No Treatment 500 mM HOCl 17C (int.std.) 19C (int.std.) 17C (int.std.) 19C (int.std.)

  14. Summary of reactive chlorinating species attack of plasmalogens Phagocyte Chemoattractant, Schiff base adducts Cell Death Cl O C C C H 14 29 H H 2-Chlorohexadecanal ( -Chloro-fatty aldehyde) a + H H H O C C C 29 14 +H O , Cl LPC Chlorohydrin PKA activation Gene activation? Effects on Endothelial Cells Cell Death? O 2 2 OH O C R 1 Myeloperoxidase O CH O 3 O C R 1 CH O P O (CH ) N 3 2 2 + CH O 3 CH O 3 Plasmenylcholine CH O P O (CH ) N 3 2 2 (Plasmalogen) + CH O 3 Lysophosphatidylcholine

  15. ACKNOWLEDGEMENTS NIH and American Heart Association Saint Louis University: Carolyn Albert Arun Thukkani Kristin Wildsmith Maria Messner Viral Brahmbhatt Dhanam Anbukumar George Vogler Brad Martinson Washington University Mass Spectrometry Resource: John Turk Fong Hsu Jan Crowley

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