title: Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
authors.forename: Hu
authors.initials: H
authors.lastname: Tao
authors.forename: Wenjun
authors.initials: W
authors.lastname: Liu
authors.forename: Brandi N
authors.initials: BN
authors.lastname: Simmons
authors.forename: Helen K
authors.initials: HK
authors.lastname: Harris
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Cox
authors.forename: Michael A
authors.initials: MA
authors.lastname: Massiah
mesh_terms: 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
chemical_list: 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