scholarly journals The use of valinomycin, nigericin and trichlorocarbanilide in control of the protonmotive force in Escherichia coli cells

1983 ◽  
Vol 212 (1) ◽  
pp. 105-112 ◽  
Author(s):  
S Ahmed ◽  
I R Booth
Keyword(s):  
Escherichia Coli ◽  
Membrane Potential ◽  
Mode Of Action ◽  
Ph Gradient ◽  
Protonmotive Force ◽  
Modes Of Action ◽  
Escherichia Coli Cells ◽  
Low Concentrations ◽  
High K ◽  
Specific Control

Valinomycin, nigericin and trichlorocarbanilide were assessed for their ability to control the protonmotive force in Escherichia coli cells. Valinomycin, at high K+ concentrations, was found to decrease the membrane potential delta phi and indirectly to decrease the pH gradient delta pH. Nigericin was found to have two modes of action. At low concentrations (0.05-2 microM) it carried out K+/H+ exchange and decreased delta pH. At higher concentrations (50 microM) it carried out a K+-dependent transfer of H+, decreasing both delta phi and delta pH. In EDTA-treated cells only the latter mode of action was evident, whereas in a mutant sensitive to deoxycholate both types of effect were observed. Trichlorocarbanilide is proposed as an alternative to nigericin for the specific control of delta pH, and it can be used in cells not treated with EDTA.

scholarly journals Evidence for an electrogenic 3-deoxy-2-oxo-d-gluconate–proton co-transport driven by the protonmotive force in Escherichia coli K12

1977 ◽  
Vol 168 (2) ◽  
pp. 211-221 ◽  
Author(s):  
A Lagarde
Keyword(s):  
Escherichia Coli ◽  
Membrane Potential ◽  
Membrane Vesicles ◽  
Ph Gradient ◽  
Protonmotive Force ◽  
Chemical Gradient ◽  
Escherichia Coli K12 ◽  
Relative Contribution ◽  
Isolated Membrane Vesicles ◽  
External Ph

Evidence is presented indicating that the carrier-mediated uptake of 3-deoxy-2-oxo-D-gluconate and D-glucuronate in Escherichia coli K12 is driven by the deltapH and deltapsi components of the protonmotive force. 1. Approximately two protons enter the cells with each sugar molecule, independent of the sugar and the strain used. 2. In respiring cells, the magnitude of the pH gradient alone, as measured by distribution of [3H]acetate, appears to be insufficient to account for the chemical gradient of 3-deoxy-2-oxo-D-gluconate that is developed between pH 6.0 and 8.0. 3. If the external pH is varied between 5.5 and 8.0, 3-deoxy-2-oxo-D-gluconate uptake is gradually inhibited by valinomycin plus K+ ions, whereas the inhibition caused by nigericin is concomitantly relieved, thus reflecting the relative contribution of deltapH and deltapsi to the total protonmotive force at each external pH. 4. 3-Deoxy-2-oxo-D-gluconate can be transiently accumulated into isolated membrane vesicles in response to an artificially induced pH gradient. The process is stimulated when the membrane potential is collapsed by valinomycin in the presence of K+ ions.

scholarly journals The effects of partial and selective reduction in the components of the proton-motive force on lactose uptake in Escherichia coli

1981 ◽  
Vol 200 (3) ◽  
pp. 583-589 ◽  
Author(s):  
S Ahmed ◽  
I R Booth
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Membrane Potential ◽  
Selective Reduction ◽  
Proton Motive Force ◽  
Motive Force ◽  
Ph Gradient ◽  
A Value ◽  
The Relationship ◽  
Lactose Transport

The relationship between the steady state lactose accumulation (delta plac) and the magnitude of the membrane potential (delta psi) and pH gradient (delta pH) has been studied at pHo5.5 and pHo7.5. An attempt has been made to differentiate between two possible means by which lactose accumulation may be reduced below the proton-motive force (delta p). Firstly, that delta psi and delta pH are not equivalent in driving lactose transport and secondly, that ‘slip’ reactions (beta-galactoside exit via the carrier without a proton) may reduce accumulation. The data support the latter; however, our conclusions are tempered by the observation that the apparent stoichiometry (delta plac/delta p) increases to a value of at least 2 at values of delta p below 130 mV.

scholarly journals Mode of action of a bactericidal protein induced in the haemolymph of Sarcophaga peregrina (flesh-fly) larvae

1984 ◽  
Vol 222 (1) ◽  
pp. 119-124 ◽  
Author(s):  
M Okada ◽  
S Natori
Keyword(s):  
Escherichia Coli ◽  
Membrane Potential ◽  
Mode Of Action ◽  
Flesh Fly ◽  
Fly Larvae

The mode of action of a bactericidal protein (sarcotoxin I) purified from the haemolymph of Sarcophaga peregrina (flesh-fly) larvae was studied, focusing attention on its effect on the function of the membrane of Escherichia coli. Sarcotoxin I almost completely blocked the uptakes of tetraphenylphosphonium ion and proline, which are known to be driven by a membrane potential, indicating that this protein disrupts the membrane potential of bacteria.

Probes of membrane potential in Escherichia coli cells

FEBS Letters ◽  
1981 ◽  
Vol 125 (2) ◽  
pp. 197-200 ◽  
Author(s):  
Alexandre Ghazi ◽  
Emanuel Schechter ◽  
Lucienne Letellier ◽  
Bernard Labedan
Keyword(s):  
Escherichia Coli ◽  
Membrane Potential ◽  
Escherichia Coli Cells

scholarly journals Different Modes of Action of Naphthyridones in Gram-Positive and Gram-Negative Bacteria

2006 ◽  
Vol 50 (1) ◽  
pp. 385-387 ◽  
Author(s):  
Ed T. Buurman ◽  
Kenneth D. Johnson ◽  
Roxanne K. Kelly ◽  
Kathy MacCormack
Keyword(s):  
Escherichia Coli ◽  
Haemophilus Influenzae ◽  
Mode Of Action ◽  
Bacterial Pathogens ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Modes Of Action

ABSTRACT Naphthyridones that were recently described as a class of translation inhibitors in gram-positive bacteria mediate their mode of action via GyrA in Haemophilus influenzae and Escherichia coli. These are the first examples of compounds in which modes of action in different bacterial pathogens are mediated through widely different targets.

scholarly journals The NADPH-oxidase-associated H+ channel is opened by arachidonate

1992 ◽  
Vol 283 (1) ◽  
pp. 171-175 ◽  
Author(s):  
L M Henderson ◽  
J B Chappell
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Membrane Potential ◽  
Nadph Oxidase ◽  
Phorbol Esters ◽  
Ph Gradient ◽  
Protonmotive Force ◽  
Phospholipid Membranes ◽  
Chemotactic Peptides ◽  
O2 Production

The H+ channel associated with the generation of O2.- by NADPH oxidase and the oxidase itself must both be activated in response to stimuli (e.g. phorbol esters, chemotactic peptides, certain fatty acids). We have investigated the effects of membrane potential, an imposed pH gradient and a combination of the two (the protonmotive force) on the H+ conductivity of the cytoplast membrane. H+ conductivity was observed only in the presence of arachidonate and not in its absence. In the presence of arachidonate, H+ movement was determined by the protonmotive force. The effect of arachidonate was probably on a channel, since this fatty acid did not significantly increase the H+ permeability of artificial phospholipid membranes. It appears, therefore, that arachidonate is required both for the activation of O2.- production and the associated H(+)-channel-mediated efflux.

scholarly journals The role of potassium transport in the generation of a pH gradient in Escherichia coli

1981 ◽  
Vol 198 (3) ◽  
pp. 691-698 ◽  
Author(s):  
R G Kroll ◽  
I R Booth
Keyword(s):  
Escherichia Coli ◽  
Membrane Potential ◽  
Respiratory Rate ◽  
Potassium Transport ◽  
Ph Gradient ◽  
Simple Relationship ◽  
K Uptake ◽  
Net Uptake ◽  
K Transport

The role of K+ transport in the generation of a pH gradient in Escherichia coli has been investigated. In K+-depleted cells, net K+ uptake dissipated delta psi (membrane potential) and led to an increase in delta pH (pH gradient). The magnitude of the delta pH formed bore a simple relationship to the net K+ uptake and was substantially independent of the respiratory rate. In K+-replete cells, generation of a pH gradient was again K+-dependent, although no net uptake of this cation occurred. The results are discussed in terms of K+ cycling, and it is suggested that delta pH is in part a function of the rate of cycling and independent of the respiratory rate.

scholarly journals Quantitative measurements of the proton-motive force and its relation to steady state lactose accumulation in Escherichia coli

1981 ◽  
Vol 200 (3) ◽  
pp. 573-581 ◽  
Author(s):  
S Ahmed ◽  
I R Booth
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Membrane Potential ◽  
Proton Motive Force ◽  
Motive Force ◽  
Ph Gradient ◽  
Experimental Conditions ◽  
Ph Values ◽  
Quantitative Measurements ◽  
Essential Prerequisite

The magnitude of delta psi (membrane potential), delta pH (pH gradient), lactose accumulation and cytoplasmic volume have been determined over a range of experimental conditions. A study of two probes of delta pH, benzoate and dimethyloxazolidene-2,4-dione (DMO), and four probes of delta psi, Rb+, K+, tetraphenylphosphonium (TPP+) and 3,3′-dipropylthiodicarbocyanine iodide, has been carried out. Benzoate and DMO are shown to be equivalent at pH values above the pK of DMO, but the latter may be less accurate below this pH. The cations TPP+ and Rb+ were found, by a number of criteria, to be equivalent, and TPP+ may be used in cells not pretreated with EDTA. These studies are an essential prerequisite to the use of TPP+ as a quantitative probe in untreated cells.

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