A circular dichroism study on thermal denaturation of a dimeric globular protein, Streptomyces subtilisin inhibitor.

Thermal denaturation of Streptomyces subtilisin inhibitor was studied by means of circular dichroism (CD) measurements in the far-UV and near-UV regions. The denaturation was found to be largely reversible; the partial irreversibility was associated with a slight loss of the inhibitory activity. Difference CD spectra in the far-UV region clarified the existence of two distinct steps in the thermal transition of the secondary structure. The first step below 80 degrees C is attributable to a partial conformational change in the alpha-helix portion, whereas the second step between 80 degrees C and 94 degrees C is attributable to a major conformational change involving the beta-sheet portion. On the assumption that the major denaturation involves dissociation of the SSI into its subunits, the enthalpy and entropy changes were determined to be 216 kcal X mol-1 and to be 603 cal X deg-1 X mol-1, respectively.