Catherine Michaux and Eric A Perpete
Unite de Chimie Physique Theorique et Structurale, University of Namur, rue de Bruxelles, 61, 5000 Namur, Belgium
Escherichia coli is one of the most widely used hosts for the production of recombinant proteins of therapeutic or commercial interest dedicated to structural and functional analysis. However, this expression system is often hampered by the formation of insoluble protein aggregates (inclusion bodies). In vitro refolding of such proteins into their native states requires screening of numerous experimental parameters specifically optimized for each system. Hence, there is currently no reliable straightforward and universal experimental solution providing the optimal refolding of proteins. The development of new original techniques in this field is therefore crucial.
In that context, we have successfully demonstrated the reliability of a new procedure for protein refolding [1-3]. This peculiar protocol is based on the association of an ionic detergent with a cosolvent. Indeed, though being known to feature denaturing abilities, some detergents appear to have their properties strongly altered when interacting with a cosolvent, and strikingly an appropriate combination of both even turns to a refolding of the protein. This remarkable procedure has successfully been applied to soluble α-helix and β-sheet peptides, as well as soluble and membrane proteins with several types of structures and properties.
In this contribution, we summarize our progress in the understanding by experimental (spectroscopy) and theoretical methods (molecular dynamics), of our recently reported approach.
Keywords: Protein refolding, detergent, cosolvent, spectroscopy, molecular dynamics.
REFERENCES:
[1] Michaux, C., Pomroy, N. C. & Privé, G. G. J. Mol. Biol. (2008) 375, 1477-1488.
[2] Roussel, G., Perpète, E. A., Matagne, A., Tinti, E. & Michaux, C. Biotechnol. Bioeng. (2013) 110, 417-423.
[3] Roussel, G., Rouse, S. L., Sansom, M. S. P., Michaux, C. & Perpète, E. A. Colloids Surfaces B Biointerfaces (2014) 114, 357-
362.