Refolding of Escherichia coli produced membrane protein inclusion bodies immobilised by nickel chelating chromatography

FEBS Letters

Kühlbrandt, W¹  Collinson, I¹  Kosemund, K²  Rogl, H¹ 

[1]Max-Planck-Institut für Biophysik, Heinrich-Hoffmann-Straße 7, D-60528 Frankfurt am Main, Germany

[2]Institut für Allgemeine Botanik der Universität, D-55099 Mainz, Germany

关键词: Inclusion body;    Refolding;    Light harvesting complex II;    Translocation pore;    Chloroplast;    C8E4;    n-octyltetraoxyethylene;    LDS;    lithium dodecylsulfate;    LDAO;    N;    N-dimethyl-dodecylamine-N-oxide;    LHC2;    light harvesting complex II;    LHCP;    LHC2 apoprotein;    OG;    octyl-glucoside;    PMSF;    phenylmethanesulfonyl fluoride;    SDS-PAGE;    sodium dodecyl sulfate polyacrylamide gel electrophoresis;    Tic;    translocon of the inner chloroplast envelope;    Toc;    translocon of the outer chloroplast envelope;   

DOI  :  10.1016/S0014-5793(98)00825-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown