Mass spectrometry evidence for formation of estrogen-homocysteine conjugates: estrogens can regulate homocysteine levels

Free Radic Biol Med. 2013 Dec:65:1447-1454. doi: 10.1016/j.freeradbiomed.2013.07.041. Epub 2013 Aug 6.

Abstract

Homocysteine (HCys), a sulfur-containing amino acid, is formed during the metabolism of methionine. An imbalance between the rate of production and the use of HCys during methionine metabolism can result in an increase in the plasma and urinary levels of HCys. HCys has been shown to be toxic to vascular endothelial cells through several pathways. Many earlier clinical studies have revealed an association between plasma HCys and cardiovascular and other diseases. In contrast, estrogens are suggested to lower the risk of cardiovascular disease. Several studies indicate that estrogen metabolites could be responsible for cardiovascular protection. It has been demonstrated that electrophilic estrogen quinones, E1(E2)-2,3-Q and E1(E2)-3,4-Q, can alkylate DNA as well as form conjugates with glutathione. I hypothesize that estrogen quinones generated in situ by oxidative enzymes, metal ions, or molecular oxygen can interact with HCys to form conjugates. This in turn could lower the levels of toxic HCys as well as quenching the reactive estrogen quinones, resulting in cardiovascular protective effects. To test the feasibility of a protective estrogen-HCys pathway, estrogen quinones were treated with HCys. Tandem mass spectrometry analysis of the assay mixture shows the formation of estrogen-HCys conjugates. Furthermore, incubation of catechol estrogens with myeloperoxidase (MPO) in the presence of HCys resulted in the formation of respective estrogen-HCys conjugates. The identities of estrogen-HCys conjugates in MPO assay extracts were confirmed by comparing them to pure synthesized estrogen-HCys standards. I propose that through conjugation estrogens could chemically regulate HCys levels; moreover these conjugates could be used as potential biomarkers in determining health.

Keywords: CVD; Cardiovascular disease; Catechol estrogens; Conjugates; E; Estrogen; Free radicals; HCys; Homocysteine; MPO; Mass spectrometry; Myeloperoxidase; Quinone.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alkylation
  • Cardiotonic Agents / chemistry
  • Cardiovascular Diseases
  • DNA / chemistry
  • Estradiol Congeners / chemical synthesis*
  • Estradiol Congeners / chemistry
  • Estrogens / chemistry*
  • Estrogens, Catechol / chemistry*
  • Glutathione / chemistry
  • Homocysteine / biosynthesis
  • Homocysteine / blood
  • Homocysteine / chemistry*
  • Mass Spectrometry
  • Oxidation-Reduction
  • Peroxidase / metabolism
  • Quinones / chemistry*

Substances

  • Cardiotonic Agents
  • Estradiol Congeners
  • Estrogens
  • Estrogens, Catechol
  • Quinones
  • Homocysteine
  • DNA
  • Peroxidase
  • Glutathione