title: Consequences of chemical modifications on the free radical reactions of human hemoglobin.
abstract: Hemoglobin-based oxygen carriers (HBOCs) are candidates for use as blood substitutes and resuscitation fluids. We determined that HBOCs of specific types differ in their ability to generate or interact with free radicals. The differences do not correlate with oxygen affinity. Detailed comparisons with unmodified human hemoglobin, HbA0, were carried out with two cross-linked derivatives: HbA-FMDA, produced by the reaction of human oxyhemoglobin with fumaryl monodibromoaspirin, and HbA-DBBF, produced by the reaction of human deoxyhemoglobin with bis(3,5-dibromosalicyl) fumarate. Both derivatives had lower oxygen affinity than unmodified HbA0. As previously reported, exposure of oxyhemoglobin to H2O2 causes generation of free radicals capable of generating formaldehyde from dimethyl sulfoxide. Relative to the reaction catalyzed by 50 microM HbA (18.0 +/- 3.5 nmol/30 min/ml), the formaldehyde formation was roughly 70% for HbA-DBBF and 50% for HbA-FMDA under comparable conditions. More profound differences are exhibited at lower hemoglobin concentrations. Spectral changes of the HBOCs during the reaction differ qualitatively and occur at different rates. The HBOCs also differ in rates of hemoglobin-catalyzed NADPH oxidation and aniline hydroxylation, reactions mediated by reactive oxygen species. These results show that stereochemical differences brought about by chemical cross-linking alter the ability of HBOCs to generate radicals and to react with activated oxygen species. These studies also show that the ability of hemoglobin to produce activated species of oxygen can be enhanced or suppressed independently of oxygen affinity.
authors.affiliation: Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892.
authors.forename: A I
authors.initials: AI
authors.lastname: Alayash
authors.forename: J C
authors.initials: JC
authors.lastname: Fratantoni
authors.forename: C
authors.initials: C
authors.lastname: Bonaventura
authors.forename: J
authors.initials: J
authors.lastname: Bonaventura
authors.forename: E
authors.initials: E
authors.lastname: Bucci
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry; D001802:Blood Substitutes / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D005609:Free Radicals; D006441:Hemoglobin A / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D008706:Methemoglobin / Q000737:chemistry; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010108:Oxyhemoglobins / Q000737:chemistry; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship; D013482:Superoxide Dismutase / Q000494:pharmacology
chemical_list: D000814:Aniline Compounds; D001802:Blood Substitutes; D003432:Cross-Linking Reagents; D005609:Free Radicals; D010108:Oxyhemoglobins; D005557:Formaldehyde; D009249:NADP; D008706:Methemoglobin; D006441:Hemoglobin A; D013482:Superoxide Dismutase; C023650:aniline