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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acyltransferases / metabolism
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Animals
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Bacterial Outer Membrane Proteins / isolation & purification
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Bacterial Outer Membrane Proteins / metabolism
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Bacteriorhodopsins / chemistry
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Buffers
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Carbonic Anhydrase II / chemistry
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Chickens
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Dose-Response Relationship, Drug
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Escherichia coli Proteins / metabolism
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Glycols / chemistry
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Halobacterium salinarum / chemistry
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Hot Temperature
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Humans
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Hydrophobic and Hydrophilic Interactions
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Inclusion Bodies / chemistry
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Muramidase / chemistry
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Protein Conformation / drug effects
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Protein Denaturation*
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Protein Folding
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Protein Renaturation*
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Protein Structure, Secondary
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Sodium Dodecyl Sulfate / pharmacology*
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Solubility
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Solvents / chemistry*
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Surface-Active Agents / pharmacology*
Substances
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Bacterial Outer Membrane Proteins
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Buffers
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Escherichia coli Proteins
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Glycols
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Solvents
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Surface-Active Agents
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Sodium Dodecyl Sulfate
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Bacteriorhodopsins
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Acyltransferases
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PagP protein, E coli
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Muramidase
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Carbonic Anhydrase II
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hexylene glycol