Role of ionic liquids in protein refolding: native/fibrillar versus treated lysozyme

Several ionic liquids (ILs) are known to revert aggregation processes and to improve the in vitro refolding of denatured/fibrillar proteins. Here, Raman spectroscopy is exploited to verify the refolding capability of several ammonium-based ILs and to identify the microscopic signatures of the structural rearrangements induced by the interaction of ILs with fibrillar lysozyme. We collected and carefully analyzed spectra from native, fibrillar and ILs-treated Several ionic liquids (ILs) are known to revert aggregation processes and to improve the in vitro refolding of denatured/fibrillar proteins. Here, Raman spectroscopy is exploited to verify the refolding capability of several ammonium-based ILs and to identify the microscopic signatures of the structural rearrangements induced by the interaction of ILs with fibrillar lysozyme. We collected and carefully analyzed spectra from native, fibrillar and ILs-treated fibrillar lysozyme to follow the microscopic process induced by ILs. These allowed us to identify different mechanisms of interaction depending on the length of the cation alkyl chain. A clear refolding effect was observed with EAN, as well as a tendency of the longer alkyl chain (PAN and BAN) of dissolving the fibril packing. A specific interaction mainly affecting the aromatic residues was identified for MEOAN (a long chain ILs with an ether group). The whole of the results, thus, provides new and detailed information on the ILs–protein interaction and shows Raman spectroscopy as a simple, reliable and effective diagnostic technique in this field.to follow the microscopic process induced by ILs. These allowed us to identify different mechanisms of interaction depending on the length of the cation alkyl chain. A clear refolding effect was observed with EAN, as well as a tendency of the longer alkyl chain (PAN and BAN) of dissolving the fibril packing. A specific interaction mainly affecting the aromatic residues was identified for MEOAN (a long chain ILs with an ether group). The whole of the results, thus, provides new and detailed information on the ILs–protein interaction and shows Raman spectroscopy as a simple, reliable and effective diagnostic technique in this field.