title: Disruption of heme-peptide covalent cross-linking in mammalian peroxidases by hypochlorous acid.
abstract: Myeloperoxidase (MPO), lactoperoxidase (LPO) and eosinophil peroxidase (EPO) play a central role in oxidative damage in inflammatory disorders by utilizing hydrogen peroxide and halides/pseudo halides to generate the corresponding hypohalous acid. The catalytic sites of these enzymes contain a covalently modified heme group, which is tethered to the polypeptide chain at two ester linkages via the methyl group (MPO, EPO and LPO) and one sulfonium bond via the vinyl group (MPO only). Covalent cross-linking of the catalytic site heme to the polypeptide chain in peroxidases is thought to play a protective role, since it renders the heme moiety less susceptible to the oxidants generated by these enzymes. Mass-spectrometric analysis revealed the following possible pathways by which hypochlorous acid (HOCl) disrupts the heme-protein cross-linking: (1) the methyl-ester bond is cleaved to form an alcohol; (2) the alcohol group undergoes an oxygen elimination reaction via the formation of an aldehyde intermediate or undergoes a demethylation reaction to lose the terminal CH2 group; and (3) the oxidative cleavage of the vinyl-sulfonium linkage. Once the heme moiety is released it undergoes cleavage at the carbon-methyne bridge either along the δ-β or a α-γ axis to form different pyrrole derivatives. These results indicate that covalent cross-linking is not enough to protect the enzymes from HOCl mediated heme destruction and free iron release. Thus, the interactions of mammalian peroxidases with HOCl modulates their activity and sets a stage for initiation of the Fenton reaction, further perpetuating oxidative damage at sites of inflammation.
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; Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA. Electronic address: habusoud@med.wayne.edu.
authors.forename: Husam M
authors.initials: HM
authors.lastname: Abu-Soud
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.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: Faten
authors.initials: F
authors.lastname: Shaeib
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: Sana N
authors.initials: SN
authors.lastname: Khan
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
authors.forename: Jaeman
authors.initials: J
authors.lastname: Byun
authors.affiliation: Department of Pediatrics, Children's Hospital of Michigan, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Abdulhamid
authors.affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Yang
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: Ghassan M
authors.initials: GM
authors.lastname: Saed
authors.affiliation: Department of Obstetrics and Gynecology, Georgia Regents University, Augusta, GA 30912, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Diamond
authors.affiliation: The University of Toledo, Department of Chemistry and School of Green Chemistry and Engineering, 2801 W. Bancroft St., Toledo, OH 43606, USA.
authors.forename: Peter R
authors.initials: PR
authors.lastname: Andreana
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA. Electronic address: spennath@med.umich.edu.
authors.forename: Subramaniam
authors.initials: S
authors.lastname: Pennathur
mesh_terms: D055162:Biocatalysis; D002853:Chromatography, Liquid; D006418:Heme / Q000737:chemistry*; D006801:Humans; D006997:Hypochlorous Acid / Q000737:chemistry*; D018384:Oxidative Stress; D010455:Peptides / Q000737:chemistry*; D010544:Peroxidases / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010455:Peptides; D006418:Heme; D006997:Hypochlorous Acid; D010544:Peroxidases
keywords: Heme destruction; Hypochlorous acid; Inflammation; Mammalian peroxidase; Oxidative stress