Polyol-induced molten globule of cytochrome c: an evidence for stabilization by hydrophobic interaction.
To address the contribution of hydrophobic interaction to the stability of molten globule (MG) of proteins, the effects of various polyols (ethylene glycol, glycerol, erythritol, xylitol, sorbitol, and inositol) on the structure of acid-unfolded horse cytochrome c were examined at pH 2, by means of circular dichroism (CD), partial specific volume, adiabatic compressibility, and differential scanning calorimetry (DSC). Addition of polyols induced the characteristic CD spectra of MG, the effect being enhanced with an increase in their concentration and chain length (the number of OH groups) of polyols except for ethylene glycol. The free energy change of MG formation by sorbitol was comparable with those for the salt-induced MG formation but the heat capacity change was negligibly small. The partial specific volume did not change within the experimental error but the adiabatic compressibility largely increased by MG formation. The sorbitol-induced MG showed a highly cooperative DSC thermogram with a large heat capacity change in comparison with the salt-induced one. These results demonstrate that polyols can stabilize the MG state of this protein through the enhanced hydrophobic interaction overcoming the electrostatic repulsion between charged residues. The stabilizing mechanism and structure of MG state induced by polyols were discussed in terms of the preferential solvent interactions and osmotic pressure of the medium, in comparison with the salt-induced one.
Kamiyama
T
T
Department of Materials Science, Faculty of Science, Hiroshima University, Higashi-Hiroshima, Japan.
Sadahide
Y
Y
Nogusa
Y
Y
Gekko
K
K
D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D003574:Cytochrome c Group / Q000737:chemistry*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D011108:Polymers; D011487:Protein Conformation; D013012:Sorbitol; D013816:Thermodynamics
D003574:Cytochrome c Group; D011108:Polymers; C024617:polyol; D013012:Sorbitol