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2006 | 53 | 3 | 563-568
Article title

Analysis of oxygen binding by hemoglobin on the basis of mean intrinsic thermodynamic quantities

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The binding data for oxygenation of human hemoglobin, Hb, at various temperatures and in the absence and presence of 2,3-diphosphoglycerate, DPG, and inositol hexakis phosphate, IHP, were analyzed for extraction of mean intrinsic Gibbs free energy, ΔGo, enthalpy, ΔHo, and entropy, ΔSo, of binding at various partial oxygen pressures. This method of analysis considers all the protein species present such as dimer and tetramer forms which were not considered by Imai et al. (Imai K et al., 1970, Biochim Biophys Acta 200: 189 - 196) , in their analysis which was based on Adair equation. In this regard, the values of Hill equation parameters were estimated with high precision at all points of the binding curve and used for calculation of ΔGo, ΔHo and ΔSo were also calculated by analysis of ΔGo values at various temperatures using van't Hoff equation. The results represent the enthalpic nature of the cooperativity in Hb oxygenation and the compensation effect of intrinsic entropy. The interpretation of results also to be, into account the decrease of the binding affinity of sites for oxygen in the presence of DPG and IHP without any considerable changes in the site - site interaction (extent of cooperativity). In other words, the interactions between bound ligands, organic phosphates and oxygen, are more due to a decreasing binding affinity and not to the reduction of the cooperative interaction between sites. The results also document the more heterotropic effect of IHP compared to DPG.

Physical description
  • Laboratory of Biophysical Chemistry, Department of Chemistry, Isfahan University, Isfahan, Iran
  • Laboratory of Biophysical Chemistry, Department of Chemistry, Isfahan University, Isfahan, Iran
  • Laboratory of Biophysical Chemistry, Department of Chemistry, Isfahan University, Isfahan, Iran
  • Adair GS (1925) The hemoglobin system VI. The oxygen dissociation curve of hemoglobin. J Biol Chem 63: 529-545.
  • Adair GS, Bock AV, Field H Jr (1925) The hemoglobin system I. Classification of reactions. J Biol Chem 63: 493-497.
  • Atha DH, Ackers GK (1974) Human hemoglobin as a function of pH and the extent of reaction. Biochemistry 13: 2376-2382.
  • Baldvin JM (1975) Structure and function of haemoglobin. In: Progress in Biophysics and Molecular Biology, vol 29, pp 225-320, Pergamon Press, Great Britain.
  • Bordbar AK, Saboury AA, Housaindokht MR, Moosavi Movahedi AA (1997) Statistical effects of the binding of ionic surfactant to protein. J Colloid Interface Sci 192: 415-419.
  • Brodersen R, Nielsen F, Christian JC, Andersen K (1987) Characterization of binding equilibrium data by a variety of fitted isotherms. Eur J Biochem 169: 487-495.
  • Brodersen R, Honore B, Pedersen AO, Klotz IM (1988) Binding constants for ligand-carrier complexes. Trends Pharmacol Sci 9: 252-257.
  • Chinchang W, Viprakasit V, Pung-Amritt P, Tanphaichitr VS, Yenchitsomanusb P (2005) Molecular analysis of unknown β-globin gene mutations using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique and its application in Thai families with β-thalassemias and β-globin variants. Clin Biochem 38: 987-996.
  • Hill, A.V. (1910) The heat produced by contracture and muscular tone. J Physiol 40: 389-403.
  • Imai K (1973) Analyses of oxygen equilibria of native and chemically modified human adult hemoglobins on the basis of Adair's stepwise oxygenation theory and the allosteric model of Monod, Wyman, and Changeux. Biochemistry 12: 798-808.
  • Imai K (1974) Hemoglobin Chesapeaks (92α, arginine leucine). Precise measurements and analysis of oxygen equilibrium. J Biol Chem 249: 7607-7612.
  • Imai K (1979) Thermodynamic aspects of the co-operativity in four-step oxygenation equilibria of haemoglobin. J Mol Biol 133: 233-247.
  • Imai K, Yonetani T (1974) pH Dependence of the Adair constants of human hemoglobin. J Biol Chem 250: 2227-2231.
  • Imai K, Yonetani T (1975) Thermodynamical studies of oxygen equilibrium of hemoglobin. J Biol Chem 250: 7093-7098.
  • Imai K, Morimoto H, Kotani M, Watari H, Hirata W, Kuroda M (1970) Studies on the function of abnormal hemoglobins I. An improved method for automatic measurement of the oxygen equilibrium curve of hemoglobin. Biochim Biophys Acta 200: 189-196.
  • Labergea M, Kovesia I, Yonetani T, Fidya J (2005) R-state hemoglobin bound to heterotropic effectors: models of the DPG, IHP and RSR13 binding sites. FEBS Lett 579: 627-632.
  • Nishino T, Tubb J, Emery DW (2006) Partial correction of murine β-thalassemia with a gammaretrovirus vector for human γ-globin. Blood Cells Mol Diss 37: 1-7.
  • Roughton FJW (1936) The thermo-chemistry of the oxygen-haemoglobin reaction II. Comparison of the heat as measured directly on purified hemoglobin with that calculated directly by the vant' Hoff isochor. Biochem J 30: 2117-2133.
  • Scalonia A, Ferrantia P, De Simoned G, Mamonea G, Sannoloa N, Malornia A (1999) Probing the reactivity of nucleophile residues in human 2,3-diphosphoglycerate/deoxy-hemoglobin complex by a specific chemical modifications. FEBS Lett 452: 190-194.
  • Shmukler BE, Brugnara C, Alper SL (2000) Structure and genetic polymorphism of the mouse KCC1 gene. Biochim Biophys Acta 1492: 353-361.
  • Stathopulos PB (2003) Taking the good out of the bad: lentiviral-based gene therapy of the hemoglobinopathies. Biotechnol Adv 21: 513-526.
  • Steiper ME, Wolfe ND, Karesh WB, Kilbourn AM, Bosi EJ, Ruvolo M (2006) The phylogenetic and evolutionary history of a novel alpha-globin-type gene in orangutans (Pongo pygmaeus). Infect Genet Evol 6: 277-286.
  • Tyuma I, Imai K, Shimizu K (1973) Analysis of oxygen equilibrium of hemoglobin and control mechanism of organic phosphates. Biochemistry 12: 1491-1498.
  • Tyuma I, Kamigawara Y, Imai K (1973) pH Dependence of the shape of the hemoglobin-oxygen equilibrium curve. Biochim Biophys Acta 310: 317-320.
  • Wyman J (1948) Heme proteins. Adv Protein Chem 4: 407-531.
  • Yamamoto Y, Nagaoka T (1998) A 1H NMR comparative study of human adult and fetal hemoglobin's. FEBS Lett 424: 169-172.
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