Refolding SDS-denatured proteins by the addition of amphipathic cosolvents |
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Authors: | Michaux Catherine Pomroy Neil C Privé Gilbert G |
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Institution: | 1 Division of Cancer Genomics and Proteomics, Ontario Cancer Institute, 101 College Street, Toronto, Ontario Canada, M5G 1L7 2 Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada 3 Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada |
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Abstract: | Sodium dodecyl sulfate (SDS) is a highly effective and widely used protein denaturant. We show that certain amphipathic cosolvents such as 2-methyl-2,4-pentanediol (MPD) can protect proteins from SDS denaturation, and in several cases can refold proteins from the SDS-denatured state. This cosolvent effect is observed with integral membrane proteins and soluble proteins from either the α-helical or the β-sheet structural classes. The SDS/MPD system can be used to study processes involving native protein states, and we demonstrate the reversible thermal denaturation of the outer membrane protein PagP in an SDS/MPD buffer. MPD and related cosolvents can modulate the denaturing properties of SDS, and we describe a simple and effective method to recover refolded, active protein from the SDS-denatured state. |
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Keywords: | MPD 2-methyl-2 4-pentanediol LDAO lauryldimethylamine oxide HCAII human carbonic anhydrase II DSC differential scanning calorimetry OG n-octyl-β-D-glucoside DM n-decyl-β-D-maltoside |
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