title: Differential heme release from various hemoglobin redox states and the upregulation of cellular heme oxygenase-1.
abstract: Despite advances in our understanding of the oxidative pathways mediated by free hemoglobin (Hb), the precise contribution of its highly reactive redox forms to tissue and organ toxicities remains ambiguous. Heme, a key degradation byproduct of Hb oxidation, has recently been recognized as a damage-associated molecular pattern (DAMP) molecule, able to trigger inflammatory responses. Equally damaging is the interaction of the highly redox active forms of Hb with other biological molecules. We determined the kinetics of heme loss from individual Hb redox states-ferrous (Fe(2+)), ferric (Fe(3+)), and ferryl (Fe(4+))-using two different heme receptor proteins: hemopexin (Hxp), a naturally occurring heme scavenger in plasma, and a double mutant (H64Y/V86F), apomyoglobin (ApoMb), which avidly binds heme released from Hb. We show for the first time that ferric Hb (Fe(3+)) loses heme at rates substantially higher than that of ferryl Hb (Fe(4+)). This was also supported by a higher expression of heme oxygenase-1 (HO-1) when ferric Hb was added to cultured lung alveolar epithelial cells (E10). The reported cytotoxicity of Hb may therefore be attributed to a combination of accelerated heme loss from the ferric form and protein radical formation associated with ferryl Hb. Targeted therapeutic interventions can therefore be designed to curb specific oxidative pathways of Hb in hemolytic anemias and when Hb is used as an oxygen-carrying therapeutic.
authors.affiliation: Laboratory of Biochemistry and Vascular Biology Center for Biologics Evaluation and Research Food and Drug Administration Silver Spring MD USA.
authors.forename: Tigist
authors.initials: T
authors.lastname: Kassa
authors.affiliation: Laboratory of Biochemistry and Vascular Biology Center for Biologics Evaluation and Research Food and Drug Administration Silver Spring MD USA.
authors.forename: Sirsendu
authors.initials: S
authors.lastname: Jana
authors.affiliation: Laboratory of Biochemistry and Vascular Biology Center for Biologics Evaluation and Research Food and Drug Administration Silver Spring MD USA.
authors.forename: Fantao
authors.initials: F
authors.lastname: Meng
authors.affiliation: Laboratory of Biochemistry and Vascular Biology Center for Biologics Evaluation and Research Food and Drug Administration Silver Spring MD USA.
authors.forename: Abdu I
authors.initials: AI
authors.lastname: Alayash
keywords: ferric; ferryl hemoglobin; heme; hemoglobin