Equilibrium oxygen binding to human hemoglobin cross-linked between the alpha chains by bis(3,5-dibromosalicyl) fumarate.
Oxygen equilibrium curves of human hemoglobin Ao (HbAo) and human hemoglobin cross-linked between the alpha chains (alpha alpha Hb) by bis(3,5-dibromosalicyl) fumarate were measured as a function of pH and chloride or organic phosphate concentration. Compared to HbAo, the oxygen affinity of alpha alpha Hb was lower, cooperativity was maintained, although slightly reduced, and all heterotropic effects were diminished. The major effect of alpha alpha-cross-linking appears to be a reduction of the oxygen affinity of R-state hemoglobin under all conditions. However, while the oxygen affinity of T-state alpha alpha Hb was slightly reduced at physiologic chloride concentration and in the absence of organic phosphates, KT was the same for both hemoglobins in the presence of 2,3-diphosphoglycerate (or high salt) and higher for alpha alpha Hb in the presence of inositol hexaphosphate. The reduced O2 affinity arises from smaller binding constants for both T- and R-state alpha alpha Hb rather than through stabilization of the low affinity conformation. All four Adair constants could be determined for alpha alpha Hb under most conditions, but a3 could not be resolved for HbAo without constraining a4, suggesting that the cross-link stabilizes triply ligated intermediates of hemoglobin.
Vandegriff
K D
KD
Division of Blood Research, Letterman Army Institute of Research, Presidio of San Francisco, California 94129-6800.
Medina
F
F
Marini
M A
MA
Winslow
R M
RM
D019794:2,3-Diphosphoglycerate; D000494:Allosteric Regulation; D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004163:Diphosphoglyceric Acids / Q000494:pharmacology; D006441:Hemoglobin A / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D008962:Models, Theoretical; D010108:Oxyhemoglobins / Q000378:metabolism*; D010833:Phytic Acid / Q000494:pharmacology
D003432:Cross-Linking Reagents; D004163:Diphosphoglyceric Acids; D046911:Macromolecular Substances; D010108:Oxyhemoglobins; C021630:bis(3,5-dibromosalicyl)fumarate; D019794:2,3-Diphosphoglycerate; D010833:Phytic Acid; D006441:Hemoglobin A; D001241:Aspirin