Characterisation of new intracellular membranes in Escherichia coli accompanying large scale over‐production of the b subunit of F1Fo ATP synthase

FEBS Letters

Runswick, Michael J.²  Karrasch, Simone²  Miroux, Bruno²  Walker, John E.²  de Kruijff, Ben¹  Huijbregts, Richard¹  Arechaga, Ignacio² 

[1] Centre for Biomembranes and Lipid Enzymology, Department of Biochemistry of Membranes, University of Utrecht, Padualaan 8, De Uithof, Utrecht, The Netherlands;The Medical Research Council Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge CB2 2XY, UK

关键词: Over-expression;    Membrane protein;    Intracytoplasmic membrane;    Escherichia coli;    IPTG;    isopropyl-2-D-thio-galactopyranoside;    TLC;    thin layer chromatography;    HPLC;    high performance liquid chromatography;    e.s.i.-m.s.;    electrospray ionisation-mass spectrometry;   

DOI  :  10.1016/S0014-5793(00)02054-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recombinant membrane proteins in Escherichia coli are either expressed at relatively low level in the cytoplasmic membrane or they accumulate as inclusion bodies. Here, we report that the abundant over-production of subunit b of E. coli F₁F_o ATP synthase in the mutant host strains E. coli C41(DE3) and C43(DE3) is accompanied by the proliferation of intracellular membranes without formation of inclusion bodies. Maximal levels of proliferation of intracellular membranes were observed in C43(DE3) cells over-producing subunit b. The new proliferated membranes contained all the over-expressed protein and could be recovered by a single centrifugation step. Recombinant subunit b represented up to 80% of the protein content of the membranes. The lipid:protein ratios and phospholipid compositions of the intracellular membranes differ from those of bacterial cytoplasmic membranes, and they are particularly rich in cardiolipin.

【 授权许可】

Unknown