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Post-translational Transformation of Methionine to Aspartate Is Catalyzed by Heme Iron and Driven by Peroxide: A NOVEL SUBUNIT-SPECIFIC MECHANISM IN HEMOGLOBIN

Strader, MB and Hicks, WA and Kassa, T and Singleton, E and Soman, J and Olson, JS and Weiss, MJ and Mollan, TL and Wilson, MT and Alayash, AI (2014) 'Post-translational Transformation of Methionine to Aspartate Is Catalyzed by Heme Iron and Driven by Peroxide: A NOVEL SUBUNIT-SPECIFIC MECHANISM IN HEMOGLOBIN.' The Journal of Biological Chemistry, 289 (32). 22342 - 22357. ISSN 1083-351X

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Abstract

A pathogenic V67M mutation occurs at the E11 helical position within the heme pockets of variant human fetal and adult hemoglobins (Hb). Subsequent post-translational modification of Met to Asp was reported in ? subunits of human fetal Hb Toms River (?67(E11)Val ? Met) and ? subunits of adult Hb (HbA) Bristol-Alesha (?67(E11)Val ? Met) that were associated with hemolytic anemia. Using kinetic, proteomic, and crystal structural analysis, we were able to show that the Met ? Asp transformation involves heme cycling through its oxoferryl state in the recombinant versions of both proteins. The conversion to Met and Asp enhanced the spontaneous autoxidation of the mutants relative to wild-type HbA and human fetal Hb, and the levels of Asp were elevated with increasing levels of hydrogen peroxide (H2O2). Using H218O2, we verified incorporation of 18O into the Asp carboxyl side chain confirming the role of H2O2 in the oxidation of the Met side chain. Under similar experimental conditions, there was no conversion to Asp at the ?Met(E11) position in the corresponding HbA Evans (?62(E11)Val ? Met). The crystal structures of the three recombinant Met(E11) mutants revealed similar thioether side chain orientations. However, as in the solution experiments, autoxidation of the Hb mutant crystals leads to electron density maps indicative of Asp(E11) formation in ? subunits but not in ? subunits. This novel post-translational modification highlights the nonequivalence of human Hb ?, ?, and ? subunits with respect to redox reactivity and may have direct implications to ?/? hemoglobinopathies and design of oxidatively stable Hb-based oxygen therapeutics.

Item Type: Article
Uncontrolled Keywords: Aspartate (Aspartic Acid) Heme Hemoglobin Mutant Post-translational Modification (PTM) Hemoglobinopathies Oxidation of Methionine
Subjects: Q Science > Q Science (General)
Divisions: Faculty of Science and Health > Biological Sciences, School of
Depositing User: Jim Jamieson
Date Deposited: 08 Jul 2015 10:37
Last Modified: 17 Aug 2017 17:35
URI: http://repository.essex.ac.uk/id/eprint/14166

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