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J Mol Biol. 2008 Feb 1;375(5):1477-88. Epub 2007 Nov 19.

Refolding SDS-denatured proteins by the addition of amphipathic cosolvents.

Author information

1
Division of Cancer Genomics and Proteomics, Ontario Cancer Institute, 101 College Street, Toronto, Ontario, Canada.

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 alpha-helical or the beta-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.

PMID:
18083190
DOI:
10.1016/j.jmb.2007.11.026
[Indexed for MEDLINE]

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