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Kinetics of Denaturation of Human and Chicken Hemoglobins in the Presence of Co-solvents

  • Ajloo, Davood (Institute of Biochemistry and Biophysics, University of Tehran) ;
  • Moosavi-Movahedi, Ali A. (Institute of Biochemistry and Biophysics, University of Tehran)
  • Received : 2002.12.02
  • Accepted : 2003.02.25
  • Published : 2003.07.31

Abstract

The stability of four hemoglobins (Hb) in dimer forms (low concentration) were investigated by the kinetics of denaturation. The rate constants of denaturation were obtained by variation of 280 nm absorption versus time in 10 mM Tris-HCl, 10 mM EDTA, pH 8.0 at $45^{\circ}C$ in the absence and presence of 0.5 M ethanol, dimethyl sulfoxide (DMSO), formamide, and glycerol. The results show the trend of rate constants in different co-solvents in the following order: chicken hemolysate < human hemolysate and chicken Hb D < chicken Hb A. The buried surface area was calculated for Hb samples in the absence of cosolvents. Accordingly, the trend points out that: chicken Hb D > chicken Hb A > human Hb A. These results suggest that both chicken hemolysate and chicken Hb D are relatively more stable than human and chicken Hb A, respectively. However, the denaturation rate constants of Hb in different co-solvents have designated the following order: ethanol > DMSO > formamide > glycerol. As a matter of fact, this phenomenon is an indication of an increase in the denaturation capacity (DC) and hydrophobicity, and a decrease in the surface tension of the solution in the preceding co-solvents.

Keywords

Denaturation capacity;Hemoglobin;Hydrophobicity;Rate constant;Solvent effect;Stability;Surface tension

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