The membrane ATPase of Escherichia coli

Roisin, Marie‐Paule; Képès, Adam

doi:10.1016/0005-2728(72)90214-9

Cited by 51 publications

(10 citation statements)

References 17 publications

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“…NaN, completely inhibited ATP synthesis at 10 mM. At similar concentrations, azide has been found to inhibit ATPase activity almost completely (21,27). These results suggest that the observed ATP synthesis was catalyzed by the BF,,F,.…”

Section: Resultsmentioning

confidence: 53%

Adenosine 5'-triphosphate synthesis energized by an artificially imposed membrane potential in membrane vesicles of Escherichia coli

Tsuchiya¹,

Rosen²

1976

J Bacteriol

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Adenosine 5'-triphosphate (ATP) synthesis driven by an artificially imposed membrane potential in right-side-out membrane vesicles ofEscherichia coli was investigated. Membrane vesicles prepared in the presence of adenosine diphosphate were loaded with K+ by incubation with 0.5 M potassium phosphate. Addition of valinomycin resulted in the synthesis of 0.2 to 0.3 nmol of ATP/mg of membrane protein, whereas no synthesis was observed after addition of nigericin. Addition of K+, dicyclohexylcarbodiimide, carbonylcyanide p-trifluoromethoxyphenylhydrazone, or azide to the assay buffer inhibited ATP synthesis. Adenosine diphosphate and Mg2+ were found to be required. Ca2 , which can replace Mg2+ for the hydrolytic activity of the Mg2+-adenosine triphosphatase (ATPase) (EC 3 6.1.3), could not replace Mg2+ in the synthetic reaction and, in fact, inhibited ATP synthesis even in the presence of Mg2+. Strain NR-70, a mutant lacking the Mg2+-ATPase, was unable to synthesize ATP using an artificially imposed membrane potential. Additionally, the Mg2+-ATPase was found to contain tightly bound ATP.

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Section: Resultsmentioning

confidence: 53%

Adenosine 5'-triphosphate synthesis energized by an artificially imposed membrane potential in membrane vesicles of Escherichia coli

Tsuchiya¹,

Rosen²

1976

J Bacteriol

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“…4, between 1 and 10 mM of azide the stimulation of respiration is not complete and can be increased by further addition of CCCP. It is known however, that azide completely inhibits BFI ATPase activity [13] in this range of concentration.…”

Section: Effect Of Uncouplers On Respiration Of Intact Cellsmentioning

confidence: 80%

Respiratory Control in Escherichia coli K 12

Burstein

Tiankova

Képès

1979

European Journal of Biochemistry

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Contrary to the general view point that respiratory control does not exist in Escherichia coli, we have been able to demonstrate this phenomenon in this micro-organism. E. coli cells growing aerobically on an oxidizable carbon source respire at a high rate, which cannot be increased by the addition of uncouplers of oxidative phosphorylation. Upon removal of the carbon source, the rate of respiration decreases progressively to reach, after 10 min, less than 1/20 of its previous value. Under these conditions the addition of an uncoupler increases the residual respiration by a factor of 3 to 9. This increase is due to the release of respiratory control. Release of respiratory control cannot be observed upon addition of ADP which does not penetrate across the membrane. Respiratory control can also be shown in the presence of non-limiting respiratory substrate provided that it is not a carbon source. This was shown by measuring the rate of oxidation of ascorbate and phenazine methosulfate.NADH oxidase activity of membrane vesicles obtained by high pressure disintegration was increased by a factor of about 1.5 upon the addition of uncouplers and also upon addition of ADP plus phosphate. In contrast, membrane vesicles depleted from BF1 ATPase (a soluble bacterial coupling factor presenting ATPase activity) did not exhibit respiratory control. It could be restored either by rebinding of BF1 ATPase or by addition of dicyclohexylcarbodiimide, an ATPase inhibitor.The limited respiratory rate of mitochondria during so called 'state 4' which can be increased by uncouplers or by ADP plus phosphate is attributed to respiratory control [l].The lack of respiratory stimulation by the same agents on respiring Escherichia coli led several investigators [2--41 to state that there was no respiratory control in E. coli. It appears that growing E. coli cells respire at maximal rate, presumably because ATP utilising synthetic processes continuously liberate ADP plus phosphate. The increase in respiration of starved cells observed upon addition of an actively transported lactose analogue, methyl p-Dthiogalactoside was interpreted also as a release of respiratory control by the utilization of metabolic energy [5]. In this paper, we describe conditions which allow the demonstration of respiratory control in intact E. coli without added carbon source by its release upon addition of uncouplers. That this is not due to limiting oxidizable substrate can be shown by adding excess ascorbate and phenazine methosulfate.Ahhreviations. CCCP, carbonylcyanide-p-trichloromethoxyphenylhydrazone; BF1 ATPase: soluble bacterial coupling factor F1 presenting ATPase activity.Addition of ADP, a non-penetrating solute, does not cause the same respiratory increase. We also describe a less pronounced increase of respiration, with NADH as the reducing substrate in a cell-free membrane preparation which responds to the addition of uncouplers as well as of ADP plus Pi. MATERIALS AND METHODSGrowth of Bacteria E. coli K12 strain 3300 constitutive for the lactose oper...

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“…4-6). These compounds inhibit the membrane-bound Ca2+, Mg2+-activated ATPase (12). Uptake of galactose, proline, and glutamine was inhibited by azide and DCCD suggesting that the ATPase was involved in the transport of these solutes.…”

Section: Effect Of Inhibitors and Uncouplers On Uptake Of Proline Glmentioning

confidence: 99%

Energetics of galactose, proline, and glutamine transport in a cytochrome‐deficient mutant of salmonella typhimurium

Singh¹,

Bragg²

1977

J Supramol Struct

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The effect of inhibitors and uncouplers on the osmotic shock-sensitive transport systems for glutamine and galactose (by the beta-methyl galactoside permease) was compared to their effect on the osmotic shock-resistant proline and galactose permease systems in cytochrome-deficient cells of Salmonella typhimurium SASY28. Both osmotic shock-sensitive and -resistant systems were sensitive to uncouplers and to inhibitors of the membrane-bound Ca2+, Mg2+-activated adenosine triphosphatase. This suggests that uptake by both types of systems is energized in these cells by an electrochemical gradient of protons formed by ATP hydrolysis through the ATPase.

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The membrane ATPase of Escherichia coli

Cited by 51 publications

References 17 publications

Adenosine 5'-triphosphate synthesis energized by an artificially imposed membrane potential in membrane vesicles of Escherichia coli

Adenosine 5'-triphosphate synthesis energized by an artificially imposed membrane potential in membrane vesicles of Escherichia coli

Respiratory Control in Escherichia coli K 12

Energetics of galactose, proline, and glutamine transport in a cytochrome‐deficient mutant of salmonella typhimurium

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