ATP Synthesis Catalyzed by the ATP Synthase of Escherichia coli Reconstituted into Liposomes
- 1 October 1994
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 225 (1) , 167-172
- https://doi.org/10.1111/j.1432-1033.1994.00167.x
Abstract
The H(+)-translocating F0F1-ATPase from Escherichia coli (EF0F1) was purified and reconstituted into preformed reverse-phase liposomes prepared from egg yolk phosphatidylcholine/phosphatidic acid. The EF0F1 liposomes were energized by an acid/base transition (pHout = 8.3; pHin = 5.0) and a superimposed K+/valinomycin diffusion potential ([K+]out = 100 mM; [K+]in = 0.6 mM) yielding a maximum rate (turnover number) of ATP synthesis of 27 +/- 8 mol ATP . mol EF0F1(-1) . s-1), i.e. 27 +/- 8 s-1. This reaction was inhibited by NH4Cl or by addition of the F0F1 inhibitor N,N'-dicyclohexylcarbodiimide. The rate of ATP synthesis measured as a function of the phosphate and ADP concentrations, can be described by Michaelis-Menten kinetics with a Km of 0.7 +/- 0.2 mM for phosphate ([ADP] = 200 microM) and a Km of 27 +/- 7 microM for ADP ([phosphate] = 5 mM), respectively.Keywords
This publication has 41 references indexed in Scilit:
- From uni-site to multi-site ATP synthesis in thylakoid membranesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1993
- Co-reconstitution of the H+-ATP synthase and cytochrome b-563c-554 complex from a thermophilic cyanobacteriumBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1993
- The rate of ATP‐synthesis as a function of ΔpH and Δψ catalyzed by the active, reduced H+‐ATPase from chloroplastsFEBS Letters, 1991
- Kinetics of oxidative phosphorylation in Paracoccus denitrificans. 1. Mechanism of ATP synthesis at the active site(s) of F0F1-ATPaseBiochemistry, 1990
- Estimation of the turnover number of bovine heart FoF1 complexes for ATP synthesisBiochemistry, 1988
- Characterization of the Na+‐stimulated ATPase of Propionigenium modestum as an enzyme of the F1F0 typeEuropean Journal of Biochemistry, 1987
- Comparison of ATP synthesis efficiencies in ATPase proteoliposomes of different complexitiesBioelectrochemistry and Bioenergetics, 1986
- Phosphorylation and phosphate-ATP exchange catalyzed by the ATP synthase isolated from Wolinella succinogenesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1985
- The rate of ATP synthesis by reconstituted CF0F1 liposomesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1985
- Reconstitution of the purified proton conductor (F0) of the adenosine triphosphatase complex from Escherichia coliFEBS Letters, 1980