Characterisation of new intracellular membranes in Escherichia coli accompanying large scale over-production of the b subunit of F(1)F(o) ATP synthase.

TitleCharacterisation of new intracellular membranes in Escherichia coli accompanying large scale over-production of the b subunit of F(1)F(o) ATP synthase.
Publication TypeJournal Article
Year of Publication2000
AuthorsArechaga, I, Miroux, B, Karrasch, S, Huijbregts, R, de Kruijff, B, Runswick, MJ, Walker, JE
JournalFEBS Lett
Volume482
Issue3
Pagination215-9
Date Published2000 Oct 06
ISSN0014-5793
KeywordsEscherichia coli, Intracellular Membranes, Lipids, Peptide Fragments, Phospholipids, Protein Conformation, Proton-Translocating ATPases
Abstract

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 F(1)F(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.

DOI10.1016/s0014-5793(00)02054-8
Alternate JournalFEBS Lett.
Citation Key10.1016/s0014-5793(00)02054-8
PubMed ID11024463