Refolding proteins by gel filtration chromatography

FEBS Letters

Kondoh, Akiko¹  Fisher, Robert J.¹  Clore, G.Marius²  Werner, Milton H.²  Gronenborn, Angela M.² 

[1] PRI/DynCorp, National Cancer Institute-Frederick Cancer Research and Development Center, MD 21701, USA;Laboratory of Chemical Physics, Building 5, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 132, 9000 Rockville Pike, Bethesda, MD 20892, USA

关键词: Protein refolding;    Gel filtration;    Technique;   

DOI  :  10.1016/0014-5793(94)00401-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have developed a facile means for the refolding of milligram quantities of purified proteins that employs gel filtration chromatography. We demonstrate by electrophoretic mobility shift and NMR spectroscopy that human ETS-1 protein, bovine ribonucelase A and E. coli integration host factor can be refolded into the native conformation using this technique. We have extended this strategy to the preparation of miligram quantities of macromolecular complexes suitable for structural analysis by NMR spectroscopy or X-ray crystallography. The diverse challenges to overcome in refolding these proteins illustrates the potential of this technique as a general approach for recovery of recombinant proteins produced as insoluble inclusion bodies.

【 授权许可】

Unknown