title: Melatonin attenuates hypochlorous acid-mediated heme destruction, free iron release, and protein aggregation in hemoglobin.
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.
authors.affiliation: Department of Obstetrics and Gynecology, The C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Dhiman
authors.initials: D
authors.lastname: Maitra
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Abdulhamid
authors.forename: Michael P
authors.initials: MP
authors.lastname: Diamond
authors.forename: Ghassan M
authors.initials: GM
authors.lastname: Saed
authors.forename: Husam M
authors.initials: HM
authors.lastname: Abu-Soud
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000378:metabolism*; D006454:Hemoglobins / Q000187:drug effects* / Q000378:metabolism*; D006997:Hypochlorous Acid / Q000494:pharmacology*; D007501:Iron / Q000378:metabolism*; D008550:Melatonin / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D009195:Peroxidase / Q000378:metabolism; D013053:Spectrophotometry
chemical_list: D006454:Hemoglobins; D006418:Heme; D006997:Hypochlorous Acid; D007501:Iron; D009195:Peroxidase; D008550:Melatonin