title: Unravelling the mechanisms of a protein refolding process based on the association of detergents and co-solvents.
abstract: A new technique associating the detergent Sodium Dodecyl Sulphate (SDS) and an alcohol-type co-solvent has been set up, showing an unexpected efficiency to refold several types of soluble or membrane proteins. The present contribution deepens the fundamental knowledge on the phenomena underlying this process, considering the refolding of two model peptides featuring the main protein secondary structures: α-helix and β-sheet. Their refolding was monitored by fluorescence and circular dichroism, and it turns out that: (i) 100% recovery of the folded structure is observed for both peptides, (ii) the highest the SDS concentration, the more co-solvent to be added to recover the peptides' native structures, (iii) a high alcohol concentration is required to alter the SDS denaturing properties, (iv) the co-solvent performance relies on its specific lipophilic-hydrophilic balanced character, (v) the size of the micelle formed by the detergent does not enter the process critical parameters, and (vi) increasing the salt concentration up to 1 M NaCl has a beneficial impact on the process efficiency. These mechanistic aspects will help us to improve the method and extend its application. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: Laboratory of Physical Chemistry of Biomolecules, Chemistry Department, University of Namur, 61 rue de Bruxelles, 5000, Namur, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Michaux
authors.affiliation: Laboratory of Physical Chemistry of Biomolecules, Chemistry Department, University of Namur, 61 rue de Bruxelles, 5000, Namur, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Roussel
authors.affiliation: Laboratory of Physical Chemistry of Biomolecules, Chemistry Department, University of Namur, 61 rue de Bruxelles, 5000, Namur, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Lopes-Rodrigues
authors.affiliation: Laboratory of Enzymology and Protein Folding, Centre for Protein Engineering, Institut de Chimie B6, University of Liège, 7 Place du 20 août, 4000, Liège, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Matagne
authors.affiliation: Laboratory of Physical Chemistry of Biomolecules, Chemistry Department, University of Namur, 61 rue de Bruxelles, 5000, Namur, Belgium.
authors.forename: E A
authors.initials: EA
authors.lastname: Perpète
mesh_terms: D000818:Animals; D003902:Detergents / Q000737:chemistry*; D000077270:Exenatide; D057927:Hydrophobic and Hydrophilic Interactions; D008116:Lizards; D008823:Micelles; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry*; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D011489:Protein Denaturation; D011493:Protein Kinase C / Q000737:chemistry*; D058849:Protein Refolding; D012965:Sodium Chloride / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D012995:Solubility; D012997:Solvents / Q000737:chemistry*; D014688:Venoms / Q000737:chemistry*; D000099221:Protein Kinase C zeta
chemical_list: D003902:Detergents; D000077270:Exenatide; D008823:Micelles; D010455:Peptides; D011493:Protein Kinase C; D012965:Sodium Chloride; D012967:Sodium Dodecyl Sulfate; D012997:Solvents; D014688:Venoms; D000099221:Protein Kinase C zeta
keywords: co-solvent; detergent; peptide; refolding; spectroscopy