title: Confirmation of a unique intra-dimer cooperativity in the human hemoglobin alpha(1)beta(1)half-oxygenated intermediate supports the symmetry rule model of allosteric regulation.
abstract: The contribution of the alpha(1)beta(1)half-oxygenated tetramer [alphabeta:alphaO(2)betaO(2)] (species 21) to human hemoglobin cooperativity was evaluated using cryogenic isoelectric focusing. The cooperative free energy of binding, reflecting O(2)-driven protein structure changes, was measured as (21)DeltaG(c) = 5.1 +/- 0. 3 kcal for the Zn/FeO(2) analog. For the Fe/FeCN analog, (21)DeltaG(c) was estimated as 4.0 kcal after correction for a CN ligand rearrangement artifact, demonstrating that ligand rearrangement does not invalidate previous conclusions regarding this species. In the context of the entire Hb cooperativity cascade, which includes eight intermediate species, the 21 tetramer is highly abundant relative to the other doubly-ligated species, providing strong support for the previously determined consensus partition function of O(2) binding and for the Symmetry Rule model of hemoglobin cooperativity (Ackers et al., Science 1992;255:54-63). Cooperativity of normal human hemoglobin is shown to depend on site-configuration, and not solely the number of O(2) bound, nor the occupancy of alpha vs. beta subunits. Verification of a unique contribution from the alpha(1)beta(1)doubly-oxygenated species to the equilibrium O(2) binding curve strongly reinforces the Symmetry Rule interpretation that the alpha(1)beta(1)dimer acts both as a structural and functional element in cooperative O(2) binding.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA. ackers@biochem.wustl.edu
authors.forename: G K
authors.initials: GK
authors.lastname: Ackers
authors.forename: J M
authors.initials: JM
authors.lastname: Holt
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.forename: Y
authors.initials: Y
authors.lastname: Grinkova
authors.forename: A L
authors.initials: AL
authors.lastname: Klinger
authors.forename: I
authors.initials: I
authors.lastname: Denisov
mesh_terms: D000494:Allosteric Regulation; D019281:Dimerization; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008956:Models, Chemical; D010100:Oxygen / Q000737:chemistry*; D013816:Thermodynamics; D015032:Zinc / Q000737:chemistry
chemical_list: D006454:Hemoglobins; D015032:Zinc; D010100:Oxygen