* To whom correspondence should be addressed.
Received September 30, 2016; Revision received November 13, 2016
Effects of solvent environments on protein refolding have gained significant attention due to their biotechnological and pharmaceutical applications. Recent advances have shown that a number of organic osmolytes have the unique ability to induce proper folding of several misfolded proteins and simultaneously inhibit aggregation during the process. In the present study, we investigated the effects of polyol osmolytes on the refolding of guanidinium chloride-denatured ribonuclease-A (RNase-A) and compared it with that of other osmolyte types. Measurements of enzymatic activity parameters (Km and kcat) clearly indicate that polyol-induced RNase-A folding enhanced its catalytic efficiency as compared to folding in the absence of osmolytes or in the presence of osmolytes of other types. Furthermore, structural characterization revealed that the increase in catalytic efficiency stems from conformational alterations of the polyol-induced folded protein molecules as compared to other types of osmolytes.
KEY WORDS: osmolytes, polyol, protein folding, refolding, catalytic efficiency, microenvironment