Cytochrome c is an oxidative stress-activated plasmalogenase that cleaves plasmenylcholine and plasmenylethanolamine at the sn-1 vinyl ether linkage

J Biol Chem. 2018 Jun 1;293(22):8693-8709. doi: 10.1074/jbc.RA117.001629. Epub 2018 Mar 12.


Plasmalogens are phospholipids critical for cell function and signaling that contain a vinyl ether linkage at the sn-1 position and are highly enriched in arachidonic acid (AA) at the sn-2 position. However, the enzyme(s) responsible for the cleavage of the vinyl ether linkage in plasmalogens has remained elusive. Herein, we report that cytochrome c, in the presence of either cardiolipin (CL), O2 and H2O2, or oxidized CL and O2, catalyzes the oxidation of the plasmalogen vinyl ether linkage, promoting its hydrolytic cleavage and resultant production of 2-AA-lysolipids and highly reactive α-hydroxy fatty aldehydes. Using stable isotope labeling in synergy with strategic chemical derivatizations and high-mass-accuracy MS, we deduced the chemical mechanism underlying this long sought-after reaction. Specifically, labeling with either 18O2 or H218O, but not with H218O2, resulted in M + 2 isotopologues of the α-hydroxyaldehyde, whereas reactions with both 18O2 and H218O identified the M + 4 isotopologue. Furthermore, incorporation of 18O from 18O2 was predominantly located at the α-carbon. In contrast, reactions with H218O yielded 18O linked to the aldehyde carbon. Importantly, no significant labeling of 2-AA-lysolipids with 18O2, H218O, or H218O2 was present. Intriguingly, phosphatidylinositol phosphates (PIP2 and PIP3) effectively substituted for cardiolipin. Moreover, cytochrome c released from myocardial mitochondria subjected to oxidative stress cleaved plasmenylcholine in membrane bilayers, and this was blocked with a specific mAb against cytochrome c Collectively, these results identify the first plasmalogenase in biology, reveal the production of previously unanticipated signaling lipids by cytochrome c, and present new perspectives on cellular signaling during oxidative stress.

Keywords: cardiolipin; cytochrome c; hydrogen peroxide; lipid peroxidation; lipid signaling; lysophospholipid; oxidative stress; phosphatidylinositol; plasmalogen; shotgun lipidomics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cytochromes c / chemistry
  • Cytochromes c / metabolism*
  • Horses
  • Humans
  • Hydrolases / metabolism*
  • Hydrolysis
  • Lipids / analysis
  • Male
  • Mitochondria, Heart / metabolism*
  • Oxidation-Reduction
  • Oxidative Stress*
  • Plasmalogens / metabolism*
  • Rabbits
  • Vinyl Compounds / chemistry*
  • Vinyl Compounds / metabolism


  • Lipids
  • Plasmalogens
  • Vinyl Compounds
  • choline plasmalogens
  • phosphatidal ethanolamines
  • vinyl ether
  • Cytochromes c
  • Hydrolases
  • plasmalogenase