Hemoglobin-based oxygen carriers (HBOCs): structural alterations that affect free radical generation.
We examined how changes in oxygen affinity brought about by different chemical modifications of hemoglobins affect their oxidation-reduction reactions. The three modified hemoglobins studied were HbA-FMDA, HbBv-FMDA, produced by the reaction of human or bovine oxyHb with fumaryl mono-dibromoaspirin; and HbA-DBBF, produced by the reaction of human deoxyHb with bis(3,5-dibromosalicyl) fumarate. Exposure of oxyHb to H2O2 causes generation of free radicals capable of cleaving dimethylsulfoxide (Me2SO) to produce formaldehyde (HCHO). Relative to the reaction rate for HbAo (630 +/- 130 M/min) the rates of HCHO formation were roughly 70% for HbA-DBBF, 50% for HbA-FMDA and 16% for HbBv-FMDA. Exposure to H2O2 also caused spectral changes at varied rates for the HBOCs analyzed. Although these rates were not directly correlated with the rates of free radical formation, addition of mannitol or thiourea slowed both the rate of spectral changes and HCHO formation. The relative ability of the ferric derivatives of the HBOCs to participate in free radical reactions was monitored by assays of non-enzymatic NADPH oxidation and aniline hydroxylation. HbBv-FMDA showed significantly slower rates than the other HBOCs in both assays. The observed differences between HBOCs in these assays indicate differences in their ability to generate or interact with free radicals.
Alayash
A I
AI
Center for Biologics Evaluation and Research, Bethesda, MD.
Ryan
B A
BA
Fratantoni
J C
JC
Bonaventura
J
J
Bonaventura
C
C
D000818:Animals; D001802:Blood Substitutes / Q000737:chemistry*; D002417:Cattle; D003432:Cross-Linking Reagents; D005609:Free Radicals; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010100:Oxygen; D017382:Reactive Oxygen Species / Q000737:chemistry; D013053:Spectrophotometry
D001802:Blood Substitutes; D003432:Cross-Linking Reagents; D005609:Free Radicals; D006454:Hemoglobins; D017382:Reactive Oxygen Species; D010100:Oxygen