Formation of Protein Adducts with Hydroperoxy-PE Electrophilic Cleavage Products During Ferroptosis

Details

• Personal Author:
  Amoscato AA ; Anthonymuthu T ; Bahar I ; Bayir H ; Kagan VE ; Kapralov O ; Mikulska-Ruminska K ; Shrivastava IH ; Shvedova AA ; Sparvero LJ ; Tyurin VA ; Tyurina YY ; Wenzel S
• Description:
  Ferroptosis is an iron dependent form of cell death, that is triggered by the discoordination of iron, lipids, and thiols. Its unique signature that distinguishes it from other forms of cell death is the formation and accumulation of lipid hydroperoxides, particularly oxidized forms of polyunsaturated phosphatidylethanolamines (PEs), which drives cell death. These readily undergo iron-catalyzed secondary free radical reactions leading to truncated products which retain the signature PE headgroup and which can readily react with nucleophilic moieties in proteins via their truncated electrophilic acyl chains. Using a redox lipidomics approach, we have identified oxidatively-truncated PE species (trPEox) in enzymatic and non-enzymatic model systems. Further, using a model peptide we demonstrate adduct formation with Cys as the preferred nucleophilic residue and PE(26:2) +2 oxygens, as one of the most reactive truncated PE-electrophiles produced. In cells stimulated to undergo ferroptosis we identified PE-truncated species with sn-2 truncations ranging from 5 to 9 carbons. Taking advantage of the free PE headgroup, we have developed a new technology using the lantibiotic duramycin, to enrich and identify the PE-lipoxidated proteins. Our results indicate that several dozens of proteins for each cell type, are PE-lipoxidated in HT-22, MLE, and H9c2 cells and M2 macrophages after they were induced to undergo ferroptosis. Pretreatment of cells with the strong nucleophile, 2-mercaptoethanol, prevented the formation of PE-lipoxidated proteins and blocked ferroptotic death. Finally, our docking simulations showed that the truncated PE species bound at least as good to several of the lantibiotic-identified proteins, as compared to the non-truncated parent molecule, stearoyl-arachidonoyl PE (SAPE), indicating that these oxidatively-truncated species favor/promote the formation of PEox-protein adducts. The identification of PEox-protein adducts during ferroptosis suggests that they are participants in the ferroptotic process preventable by 2-mercaptoethanol and may contribute to a point of no return in the ferroptotic death process. [Description provided by NIOSH]
• Subjects:
  Free Radicals Lipids Occupational Health Safety
• Keywords:
  2-Mercaptoethanol Author Keywords: Ferroptotic Death Cell Death Free Radicals Lantibiotics Oxidatively-truncated Lipids Oxidized Phosphatidylethanolamine Protein Biochemistry Protein Lipidation Proteins
• ISSN:
  2213-2317
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  Massachusetts ; New York ; OSHA Region 1 ; OSHA Region 2 ; OSHA Region 3 ; Pennsylvania ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  63
• NIOSHTIC Number:
  nn:20067711
• Citation:
  Redox Biol 2023 Jul; 63:102758
• Contact Point Address:
  V.E. Kagan, Center for Free Radical and Antioxidant Health, Department of Environmental and Occupational Health, University of Pittsburgh School of Public Health, 130 Desoto St, Pittsburgh, PA, 15261, USA
• Email:
  kagan@pitt.edu
• Federal Fiscal Year:
  2023
• NORA Priority Area:
  Manufacturing
• Peer Reviewed:
  True
• Source Full Name:
  Redox Biology
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:e079cd773b3504da554f42cf9e2e86dd1bac7985ee3f6bea5e785d3d90a5085611bb503189a2ad251b486f64c4acf8d5e1c43df320fa7bf8d61026fb8c0140d9
• Download URL:
  https://stacks.cdc.gov/view/cdc/229974/cdc_229974_DS1.pdf
• File Type:
  [PDF - 9.91 MB ]