Thermodynamic Study of Intermediate State of Papain Induced by n-Alkyl Sulfates at Two Different pH Values: A Spectroscopic Approach

摘要

The formation of the intermediate state of papain was induced by n-alkyl sulfates including sodium octyl sulfate, SOS; sodium decyl sulfate, SDeS; and sodium dodecyl sulfate, SDS at different concentrations. A systematic investigation of n-alkyl sulfates induced conformational alteration in molten globule state under an acidic condition and the native state of papain was examined by tryptophan fluorescence, 1-anilino 8-naphtalene sulfonic acid (ANS) binding and UV absorbance. The addition of n-alkyl sulfates to molten globule state at pH 2 shows a decrease in tryptophan fluorescence intensity and quenched ANS fluorescence relative to the native state that leads to enhancement in tryptophan fluorescence and an increase in ANS fluorescence as well. In the presence of n-alkyl sulfates in various conditions, two intermediate (IA and IB) with different conformations were obtained at acidic and native states, respectively. Thus, we can assume that the intermediate states in folding and unfolding pathways in various conditions have different structures. The results show that SDS is much more effective than SDeS and SOS for the formation of the intermediate states for papain in the two different pathways due to the presence of its hydrophobic tail. Therefore, hydrophobic interactions play an important role in inducing the two different intermediates along the two various thermodynamic pathways.