Refolding of Escherichia coli produced membrane protein inclusion bodies immobilised by nickel chelating chromatography.
Two distinctly different membrane proteins, which produced inclusion bodies in Escherichia coli, have been refolded to reconstitute properties appropriate to their native counterparts. The method employed utilises nickel chelating chromatography, where the solubilised inclusion bodies bind, refold and elute. Our aims were to release a large pool of membrane protein for functional, mutational and crystallisation screening studies. It is hoped that the methods described here will have a general application for other membrane proteins which have formed inclusion bodies.
Max-Planck-Institut für Biophysik, Frankfurt am Main, Germany.
H
H
Rogl
K
K
Kosemund
W
W
Kühlbrandt
I
I
Collinson
D002614:Chelating Agents; D002845:Chromatography / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D002479:Inclusion Bodies; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D009532:Nickel; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics; D010940:Plant Proteins; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry*
D002614:Chelating Agents; D008565:Membrane Proteins; C103933:OEP75 protein precursor, plant; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D009532:Nickel