The Interaction of Sickle Hemoglobin with DPG, CO2 and with Other Hemoglobins: Formation of Asymmetrical Hybrids

  • H. Franklin Bunn
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 28)


In either the red cell or in concentrated solution, deoxyhemoglobin S (α2β2 6 val) possesses the unique property of aggregating into microfilaments, leading to increased viscosity and ultimately to gelation. This property may be strongly influenced by the presence of non-S hemoglobins or by a second amino acid sub-stitution on either the β chain (hb C-Harlem (α2β2 6 val 73 asn) (Bookchin et. Al., 1967) or the α chain (hb Memphis (α2 23 gln β 6 val) (Kraus et al., 1966). Furthermore, the sickling phenomenon may be affected by intracellular modifiers of hemoglobin function such as carbon dioxide (CO2) and 2,3-diphosphoglycerate (DPG). Both of these co-factors bind to hemoglobin at N-terminal amino groups of globin polypeptide chains. Carbamino formation occurs at the N-terminal amino groups of both the α and the β chains (Kilmartin and Rossi-Bernardi, 1969) while DPG binds to positively charged sites on the β chains including the α amino group of NA1 valine, the imidazole of H21 histidine and the ɛ amino group of EF6 lysine (Perutz, 1970). It is of interest that sickling is inhibited by cyanate, an agent which reacts preferentially with N-terminal amino groups of proteins (Cerami and Manning, 1971).


Oxygen Equilibrium Human Hemoglobin Hybrid Molecule Bohr Effect Asymmetrical Hybrid 


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  1. Allen, D.W., Schroeder, W.A., and Balog, J. (1958). Observations on the chromatographic heterogeneity of hemoglobin: A study of the effects of crystallization and chromatography on the heterogeneity and isoleucine content. J. Amer. Chem. Soc. 80: 1628.CrossRefGoogle Scholar
  2. Bauer, C. (1969). Antagonistic influence of CO2 and 2,3-diphosphoglycerate on the Bohr effect of human haemoglobin. Life Sci. 8: 1041.PubMedCrossRefGoogle Scholar
  3. Benesch, R.E., Benesch, R., and Xu, C.I. (1969). The oxygenation of hemoglobin in the presence of 2,3-diphosphoglycerate. Effect of temperature, pH, ionic strength and hemoglobin con-centration. Biochemistry 8: 2567.PubMedCrossRefGoogle Scholar
  4. Bertles, J.F., Rabinowitz, R., and Döbler, J. (1970). Hemoglobin interaction: modification of solid phase composition in the sickling phenomenon. Science 169: 375.PubMedCrossRefGoogle Scholar
  5. Beutler, E., Paniker, N.V., and West, C. (1971). The effect of 2,3-DPG on the sickling phenomenon. Blood 37: 184.PubMedGoogle Scholar
  6. Bunn, H.F., and Briehl, R.W. (1970). The interaction of 2,3-diphosphoglycerate with various human hemoglobins. J. Clin. Invest. 49: 1088.PubMedCrossRefGoogle Scholar
  7. Bunn, H.F., Bradley, T.B., Davis, W.E., Drysdale, J.W., Burke, J.F., Beck, F.S., and Laver, M.B. (1972). Structural and functional studies on hemoglobin Bethesda (α2β2 145 His), a variant associated with compensatory erythrocytosis. J. Clin. Invest. In press.Google Scholar
  8. Cerami, A., and Manning, J.M. (l97l). Potassium cyanate as an in-hibitor of the sickling of red blood cells in vitro. Proc. Hat. Acad. Sci. U.S.A. 68:1180.CrossRefGoogle Scholar
  9. Drabkin, D. (1946). Spectrophotometry studies XIY. The crystallo-graphic and optical properties of the hemoglobin of man in comparison with those of other species. J. Biol. Chem. 164:703PubMedGoogle Scholar
  10. Drysdale, J.W., Righetti, P., and Bunn, H.F. (1971). The separa-tion of human and animal hemoglobins by isoelectric focusing on Polyacrylamide gel. Biochim. Biophys. Acta 229: 42.Google Scholar
  11. Edelstein, S.J., Rehmar, M.J., Olson, J.S., and Gibson, Q.H. (1970). Functional aspects of the subunit association-dissociation equilibria of hemoglobin. J. Biol. Chem. 245: 4372.PubMedGoogle Scholar
  12. Guidotti, G., Königsberg, W.H., and Craig, L.C. (1963). On the dissociation of normal adult human hemoglobin. Proc. Nat. Acad. Sci. U.S.A. 50: 774.CrossRefGoogle Scholar
  13. Jensen, M., Bunn, H.F., Halikas, G., and Nathan, D. (1972). Oxygen-affinity dependent and independent inhibition of sickling by cyanate. Second International Conference on Red Cell Metabolism and Function, G. Brewer, editor, Plenum Press, New YorkGoogle Scholar
  14. Kilmartin, J.Y., and Rossi-Bernardi, L. (1969). Inhibition of CO2 combination and reduction of the Bohr effect in haemoglobin chemically modified at its α-amino groups. Nature 222: 1243.PubMedCrossRefGoogle Scholar
  15. Kraus, L.M., Miyaji, T., Iuchi, I., and Kraus, A.P. (1966). Characterization of α23 glu NH2 in hemoglobin Memphis. Hemoglobin Memphis/S, a new variant of molecular disease. Biochemistry 5: 3701.PubMedCrossRefGoogle Scholar
  16. Lian, C.Y., Both, S., and Harkness, D.R. (1971). The effect of alteration of intracellular 2,3-DPG- concentration upon oxygen “binding of intact erythrocytes containing normal and mutant hemoglobins”. Biochem. Biophys. Res. Commun. 45: 151.PubMedCrossRefGoogle Scholar
  17. Park, C.M. (1970). The dimerization of deoxyhemoglobin and of oxyhemoglobin. J. Biol. Chem. 245: 5390.PubMedGoogle Scholar
  18. Perutz., M.F. (1970). Stereochemistry of cooperative effects in Haemoglobin. Nature 228: 726.PubMedCrossRefGoogle Scholar
  19. Rossi-Bernardi, L., Houghton, F.J.W., Pace, M., and Coven, E. (1972). The effect of organic phosphates on the binding of CO2 to human hemoglobin and CO2 transport in the circulating blood. In Oxygen Affinity of Hemoglobin and led Cell Acid-Base Status, M. Rorth and P. Astrup, editors, p. 225 9 Munksgaard, Copenhagen.Google Scholar
  20. Tomita, S., and Riggs, A. (1971). Studies of the interaction of 2,3-diphosphoglycerate and carbon dioxide with hemoglobins from man, mouse and elephant. J. Biol. Chem. 246: 547.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1972

Authors and Affiliations

  • H. Franklin Bunn
    • 1
  1. 1.Thorndike Memorial Laboratory, Boston City HospitalHarvard Medical SchoolBostonUSA

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