Refolding SDS-denatured proteins by the addition of amphipathic cosolvents.
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.
Michaux
Catherine
C
Division of Cancer Genomics and Proteomics, Ontario Cancer Institute, 101 College Street, Toronto, Ontario, Canada.
Pomroy
Neil C
NC
Privé
Gilbert G
GG
D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000737:chemistry; D002021:Buffers; D024402:Carbonic Anhydrase II / Q000737:chemistry; D002645:Chickens; D004305:Dose-Response Relationship, Drug; D029968:Escherichia coli Proteins / Q000378:metabolism; D006018:Glycols / Q000737:chemistry; D019617:Halobacterium salinarum / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D002479:Inclusion Bodies / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D017433:Protein Structure, Secondary; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D012995:Solubility; D012997:Solvents / Q000737:chemistry*; D013501:Surface-Active Agents / Q000494:pharmacology*
D001425:Bacterial Outer Membrane Proteins; D002021:Buffers; D029968:Escherichia coli Proteins; D006018:Glycols; D012997:Solvents; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate; D001436:Bacteriorhodopsins; D000217:Acyltransferases; C468520:PagP protein, E coli; D009113:Muramidase; D024402:Carbonic Anhydrase II; C011480:hexylene glycol