Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS.
We describe a rapid, simple, and efficient method for recovering glutathione S-transferase (GST)- and His6-tagged maltose binding protein (MBP) fusion proteins from inclusion bodies. Incubation of inclusion bodies with 10% sarkosyl effectively solubilized >95% of proteins, while high-yield recovery of sarkosyl-solubilized fusion proteins was obtained with a specific ratio of Triton X-100 and CHAPS. We demonstrate for the first time that this combination of three detergents significantly improves binding efficiency of GST and GST fusion proteins to gluthathione (GSH) Sepharose.
Tao
Hu
H
Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
Liu
Wenjun
W
Simmons
Brandi N
BN
Harris
Helen K
HK
Cox
Timothy C
TC
Massiah
Michael A
MA
D002793:Cholic Acids; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D002479:Inclusion Bodies / Q000737:chemistry*; D058619:Maltose-Binding Proteins; D017830:Octoxynol; D033902:Periplasmic Binding Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D012521:Sarcosine / Q000031:analogs & derivatives; D012995:Solubility
D002793:Cholic Acids; D058619:Maltose-Binding Proteins; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; C025231:sarkosyl; D017830:Octoxynol; D005982:Glutathione Transferase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; D012521:Sarcosine