title: Method for quantitative refolding of the link module from human TSG-6.
abstract: We have developed a procedure for the quantitative refolding of the Link module from human tumor necrosis factor-stimulated gene 6. This significantly simplifies the previously described method of production of this protein domain (Day et al., Protein Expression Purif. 8, 1-16, 1996). The refolding is carried out under nondenaturing conditions at pH 6.0 in the presence of a 100-fold molar excess of beta-mercaptoethanol. After 2 days the starting material, which consists of three species that differ only with respect to their disulfide bond organization, has rearranged to give a single homogeneous species with the correct disulfide bridges. This method allows the production of about 20 mg of folded protein per liter of Escherichia coli culture.
authors.affiliation: Department of Biochemistry, University of Oxford, United Kingdom.
authors.forename: J D
authors.initials: JD
authors.lastname: Kahmann
authors.forename: R
authors.initials: R
authors.lastname: Koruth
authors.forename: A J
authors.initials: AJ
authors.lastname: Day
mesh_terms: D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D015815:Cell Adhesion Molecules; D004220:Disulfides; D010446:Peptide Fragments; D011994:Recombinant Proteins; C072273:TNFAIP6 protein, human; D003545:Cysteine