Refolding SDS-mediated partially-unfolded proteins: A distinguishing role of tetraalkylammonium ionic liquids via mixed micellar aggregation

Abstract

The current study reports a new strategy for ionic liquid (ILs) assisted refolding of sodium dodecyl sulfate (SDS)-induced partially unfolded protein via mixed micellar aggregate formation between SDS and the ionic liquid. This approach can indeed be utilized to keep many essential and sensitive proteins as active and prolong their shelf life. For SDS-induced partially unfolded proteins, the ionic liquid has been identified to function as a refolding agent. The phenomenon has been observed by us for the first time and we have thoroughly investigated the same utilizing a variety of spectroscopic, microscopic approaches, and attempted to rationalize from computational techniques as well. The steady-state fluorescence emission spectra and time-resolved fluorescence decay show that both the intrinsic emission intensity and the average lifetime, respectively, of the surfactant-induced partially-unfolded protein increase in the presence of ionic liquid, indicating a possible reversal of unfolding (refolding) of the protein (up to ∼95 % revival of luminescence intensity and average lifetime with respect to the native protein). The CD and FTIR results also confirmed that the α-helical content of the pre-unfolded protein reverts substantially and is very close to that of the native protein. To understand the refolding process of SDS-induced partially-unfolded protein, we have further studied the structural recovery of urea-induced pre-denatured protein utilizing the same ionic liquid. Interestingly, no such refolding was observed in this case. From the experimental results, it appears that ionic liquid is involved in scavenging the unfolding surfactant monomers by forming mixed-micellar aggregates where the ion-ion attachment is likely to have impact on the said process.