title: Effectors of hemoglobin. Separation of allosteric and affinity factors.
abstract: The relative contributions of the allosteric and affinity factors toward the change in p50 have been calculated for a series of effectors of hemoglobin (Hb). Shifts in the ligand affinity of deoxy Hb and the values for 50% ligand saturation (p50) were obtained from oxygen equilibrium data. Because the high-affinity parameters (liganded conformation) are poorly determined from the equilibrium curves, they were determined from kinetic measurements of the association and dissociation rates with CO as ligand. The CO on-rates were obtained by flash photolysis measurements. The off-rates were determined from the rate of oxidation of HbCO by ferricyanide, or by replacement of CO with NO. The partition function of fully liganded hemoglobin for oxygen and CO is only slightly changed by the effectors. Measurements were made in the presence of the effectors 2,3-diphosphoglycerate (DPG), inositol hexakisphosphate (IHP), bezafibrate (Bzf), and two recently synthesized derivatives of Bzf (LR16 and L35). Values of p50 change by over a factor of 60; the on-rates decrease by nearly a factor of 8, with little change in the off-rates for the liganded conformation. The data indicate that both allosteric and affinity parameters are changed by the effectors; the changes in ligand affinity represent the larger contribution toward shifts in p50.
authors.affiliation: INSERM U299, Hôpital de Bicêtre, France.
authors.forename: M C
authors.initials: MC
authors.lastname: Marden
authors.forename: B
authors.initials: B
authors.lastname: Bohn
authors.forename: J
authors.initials: J
authors.lastname: Kister
authors.forename: C
authors.initials: C
authors.lastname: Poyart
mesh_terms: D000495:Allosteric Site; D001665:Binding Sites; D002263:Carboxyhemoglobin / Q000378:metabolism; D006441:Hemoglobin A / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008962:Models, Theoretical; D009569:Nitric Oxide / Q000097:blood; D010108:Oxyhemoglobins / Q000378:metabolism*; D010782:Photolysis
chemical_list: D010108:Oxyhemoglobins; D009569:Nitric Oxide; D006441:Hemoglobin A; D002263:Carboxyhemoglobin; C029774:oxyhemoglobin A