Control of membrane potential by external H+ concentration in Bacillus subtilis as determined by an ion-selective electrode

Hosoi, Shigeru; Mochizuki, Noriko; Hayashi, Shigeru; Kasai, Michiki

doi:10.1016/0005-2736(80)90487-3

Cited by 15 publications

(7 citation statements)

References 18 publications

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“…Using this approach, we estimated the membrane potential of B. subtilis to reach ~ −110 mV under the growth conditions used in our experiments. This value is in good agreement with previously published estimates (Hosoi et al, 1980 ; Zaritsky et al, 1981 ).…”

Section: Resultssupporting

confidence: 93%

Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

Winkel

Gray

Seistrup

et al. 2016

Front. Cell Dev. Biol.

242

289

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The bacterial cytoplasmic membrane is a major inhibitory target for antimicrobial compounds. Commonly, although not exclusively, these compounds unfold their antimicrobial activity by disrupting the essential barrier function of the cell membrane. As a consequence, membrane permeability assays are central for mode of action studies analysing membrane-targeting antimicrobial compounds. The most frequently used in vivo methods detect changes in membrane permeability by following internalization of normally membrane impermeable and relatively large fluorescent dyes. Unfortunately, these assays are not sensitive to changes in membrane ion permeability which are sufficient to inhibit and kill bacteria by membrane depolarization. In this manuscript, we provide experimental advice how membrane potential, and its changes triggered by membrane-targeting antimicrobials can be accurately assessed in vivo. Optimized protocols are provided for both qualitative and quantitative kinetic measurements of membrane potential. At last, single cell analyses using voltage-sensitive dyes in combination with fluorescence microscopy are introduced and discussed.

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Section: Resultssupporting

confidence: 93%

Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

Winkel

Gray

Seistrup

et al. 2016

Front. Cell Dev. Biol.

242

289

View full text Add to dashboard Cite

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“…[methoxy-14C]inulin (0.05 mCi/2.8 mg) was obtained from New England Nuclear Corp. TPP+ was purchased from Merck (Germany), and carbonyl cyanide-m-chlorophenyl hydrazone (CCCP) was a product of Tokyo Kasei Co. (Japan). Other chemicals used were described previously (9,28).…”

Section: Materlals and Methodsmentioning

confidence: 99%

“…Cell concentration was adjusted to an absorbancy at 590 nm of 4.0. TPP+ uptake by the cells was monitored with a TPP+-selective electrode at 30°C, and the AO\ was calculated as described previously (9).…”

Section: Measurement Of ['H]tpmp+ Uptake [3h]-mentioning

confidence: 99%

Use of lipophilic cation-permeable mutants for measurement of transmembrane electrical potential in metabolizing cells of Escherichia coli

Hirota¹,

Matsuura²,

Mochizuki³

et al. 1981

J Bacteriol

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Some lipopolysaccharide-defective mutants of Escherichia coli showed, without ethylenediaminetetraacetic acid treatment, a quick and high uptake of lipophilic cations such as triphenylmethylphosphonium and tetraphenylphosphonium. The rate and amount of uptake were comparable to those of an ethylenediaminetetraacetic acid-treated wild type. Transmembrane electrical potential, which was calculated from the distribution of these lipophilic cations between the inside and outside of the mutant cells, was about -150 mV at pH 7.5 and showed a strong dependency on the external pH. One of the E. coli mutants, the acrA mutant, was found to be also permeable to dicyclohexylcarbodiimide, an H+-adenosine triphosphatase inhibitor, and 1-anilino-8-naphthalene sulfonate, a fluorescent dye. The acrA mutant was vigorously motile and highly sensitive to many bacteriophages and colicins. Thus, the acrA mutant is quite useful for the quantitative measurement of transmembrane electrical potential by lipophilic cations in intact and metabolizing cells especially in relation to motility and actions of colicins and bacteriophages.

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“…Bacteria were added to an OD 595 of 4.0 in 10 mL of the same buffer supplemented with glucose and TPP + to a concentration of 5 mM and 10 mM, respectively, and TPP + uptake was measured with the TPP + -selective electrode method described by Hosoi et al 46 TPP + concentration in the external medium was determined with a Kwik-Tip Ag/AgCl half-cell electrode, which was constructed according to the instructions provided by the manufacturer (World Precision Instruments, Sarasota, FL, USA). Both the TPP + -selective electrode and a Flexible Dri-Ref reference electrode (World Precision Instruments, Sarasota, FL, USA) were connected to a Jenway 3510 ion meter (Cole-Parmer, Staffordshire, UK) and the LabTrax 4-Channel Data Acquisition system (WPI).…”

Section: Qrt-pcrmentioning

confidence: 99%

Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli

Frimodt-Møller

Koulouktsis

Charbon

et al. 2021

Molecular Therapy - Nucleic Acids

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Cell-penetrating peptides (CPPs) are increasingly used for cellular drug delivery in both pro-and eukaryotic cells, and oligoarginines have attracted special attention. How arginine-rich CPPs translocate across the cell envelope, particularly for prokaryotes, is still unknown. Arginine-rich CPPs efficiently deliver antimicrobial peptide nucleic acid (PNA) to its intracellular mRNA target in bacteria. We show that resistance to PNA conjugated to an arginine-rich CPP in Escherichia coli requires multiple genetic modifications and is specific for the CPP part and not to the PNA part. An integral part of the resistance was the constitutively activated Cpx-envelope stress response system (cpx*), which decreased the cytoplasmic membrane potential. This indicates an indirect energy-dependent uptake mechanism for antimicrobials conjugated to arginine-rich CPPs. In agreement, cpx* mutants showed low-level resistance to aminoglycosides and an arginine-rich CPP conjugated to a peptide targeting the DNA sliding clamp, i.e., similar uptake in E. coli for these antimicrobial compounds.

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Control of membrane potential by external H+ concentration in Bacillus subtilis as determined by an ion-selective electrode

Cited by 15 publications

References 18 publications

Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

Use of lipophilic cation-permeable mutants for measurement of transmembrane electrical potential in metabolizing cells of Escherichia coli

Activating the Cpx response induces tolerance to antisense PNA delivered by an arginine-rich peptide in Escherichia coli

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