REFOLDING PROTEINS BY GEL-FILTRATION CHROMATOGRAPHY

被引:82
|
作者
WERNER, MH
CLORE, GM
GRONENBORN, AM
KONDOH, A
FISHER, RJ
机构
[1] NIDDKD,CHEM PHYS LAB,BETHESDA,MD 20892
[2] PRI DYNCORP,NCI,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21701
关键词
PROTEIN REFOLDING; GEL FILTRATION; TECHNIQUE;
D O I
10.1016/0014-5793(94)00401-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a facile means for the refolding of miligram 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.
引用
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页码:125 / 130
页数:6
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