Melatonin attenuates hypochlorous acid-mediated heme destruction, free iron release, and protein aggregation in hemoglobin

Dhiman Maitra, Ibrahim Abdulhamid, Michael Peter Diamond, Ghassan M. Saed, Husam M. Abu-Soud

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

In inflammatory diseases, where hypochlorous acid (HOCl) is elevated, iron homeostasis is disturbed, resulting in accumulation of free iron. Free iron is toxic by virtue of its ability to generate free radicals through the Fenton reaction. HOCl is generated by myeloperoxidase, (MPO) using chloride and hydrogen peroxide as substrates. Recent studies demonstrate that HOCl binds to the heme moiety of hemoglobin (Hb), which generates a transient ferric species whose formation and decay kinetics indicate it participates in protein aggregation, heme destruction, and free iron release. Here, we show that melatonin prevents HOCl-mediated Hb heme destruction and protein aggregation, using a combination of UV-vis spectrophotometry, ferrozine colorimetric assay, and in-gel heme staining. We also show that melatonin treatment prevents HOCl-mediated loss of red blood cell (RBC) viability, indicating biologic relevance of this finding. The mechanism by which melatonin prevents HOCl-mediated Hb heme destruction is by direct scavenging of HOCl and/or through the destabilization of the higher Hb oxidative states intermediates, ferryl porphyrin radical cation Hb-Fe(IV)=O+π• and Hb-Fe(IV)=O, which are formed through the reaction of HOCl with Hb. Our work establishes a direct mechanistic link between melatonin and its protective effect in chronic inflammatory diseases. Collectively, in addition to acting as an antioxidant and as a MPO inhibitor, melatonin can also exert its protective effect by inhibiting HOCl-mediated heme destruction of hemoproteins and subsequent free iron release.

Original languageEnglish (US)
Pages (from-to)198-205
Number of pages8
JournalJournal of Pineal Research
Volume53
Issue number2
DOIs
StatePublished - Sep 1 2012

Fingerprint

Hypochlorous Acid
Melatonin
Heme
Hemoglobins
Iron
Proteins
Peroxidase
Ferrozine
Poisons
Spectrophotometry
Porphyrins
Hydrogen Peroxide
Free Radicals
Cations
Chlorides
Cell Survival
Homeostasis
Chronic Disease
Antioxidants
Erythrocytes

Keywords

  • RBCs
  • free iron
  • hemoglobin
  • hypochlorous acid
  • inflammation
  • mammalian peroxidase
  • myeloperoxidase
  • oxidative stress

ASJC Scopus subject areas

  • Endocrinology

Cite this

Melatonin attenuates hypochlorous acid-mediated heme destruction, free iron release, and protein aggregation in hemoglobin. / Maitra, Dhiman; Abdulhamid, Ibrahim; Diamond, Michael Peter; Saed, Ghassan M.; Abu-Soud, Husam M.

In: Journal of Pineal Research, Vol. 53, No. 2, 01.09.2012, p. 198-205.

Research output: Contribution to journalArticle

Maitra, Dhiman ; Abdulhamid, Ibrahim ; Diamond, Michael Peter ; Saed, Ghassan M. ; Abu-Soud, Husam M. / Melatonin attenuates hypochlorous acid-mediated heme destruction, free iron release, and protein aggregation in hemoglobin. In: Journal of Pineal Research. 2012 ; Vol. 53, No. 2. pp. 198-205.
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