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. 1973 Aug;134(4):1015–1021. doi: 10.1042/bj1341015

Reconstitution of oxidative phosphorylation and the adenosine triphosphate-dependent transhydrogenase activity by a combination of membrane fractions from uncA− and uncB− mutant strains of Escherichia coli K12

G B Cox 1, F Gibson 1, L McCann 1
PMCID: PMC1177910  PMID: 4271644

Abstract

1. Membrane preparations from both uncA− and uncB− mutant strains of Escherichia coli K12, in which electron transport is uncoupled from phosphorylation, were fractionated by washing with a low-ionic-strength buffer. The fractionation gave a `5mm-Tris wash' and a `membrane residue' from each strain. This technique, applied to membranes from normal cells, separates the Mg2+,Ca2+-stimulated adenosine triphosphatase activity from the membrane-bound electron-transport chain and the non-energy-linked transhydrogenase activity. 2. Reconstitution of both oxidative phosphorylation and the ATP-dependent transhydrogenase activity was obtained by a combination of the `membrane residue' from strain AN249 (uncA−) with the `5mm-Tris wash' from strain AN283 (uncB−). 3. Valinomycin plus NH4+ inhibited oxidative phosphorylation both in membranes from a normal strain of E. coli and in the reconstituted membrane system derived from the mutant strains. 4. The electron-transport-dependent transhydrogenase activity was located in the membrane residue and was de-repressed in both the mutant strains. 5. The spatial and functional relationships between the proteins specified by the uncA and uncB genes and the transhydrogenase protein are discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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