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 β-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.
Original language | English |
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Pages (from-to) | 315-318 |
Number of pages | 3 |
Journal | Protein Expression and Purification |
Volume | 9 |
Issue number | 3 |
DOIs | |
Publication status | Published - Apr 1997 |