Site of Interaction between Phenazine Methosulphate and the Respiratory Chain of <i>Bacillus subtilis</i>

Bisschop, A.; Bergsma, Jack; Konings, Wil N.

doi:10.1111/j.1432-1033.1979.tb12832.x

Cited by 17 publications

(8 citation statements)

References 21 publications

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“…The highest efficiency (1 5 mol NADH oxidized/mol L-glutamate transported) is observed at low rates of NADH oxidation and this efficiency decreases at higher oxidation rates. Similar high efficiencies were found for NADH oxidation via membrane-bound phenazine methosulphate (9-13.4 mol NADH oxidizedjmol L-glutamate transported) [7]. With menaquinone-2 the efficiency of NADH oxidation is at low oxidation rates comparable with the efficiency observed with plumbagin but the efficiency increases to 60 (mol NADH oxidized/mol L-glutamate transported) at high N A D H oxidation rates.…”

Section: Restoration Of the Respiratory Chain Of B Subtilis Arod Witsupporting

confidence: 71%

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Restoration of NADH Oxidation with Menaquinones and Menaquinone Analogues in Membrane Vesicles from the Menaquinone‐Deficient Bacillus subtilis aroD

Bergsma¹,

Meihuizen²,

Oeveren³

et al. 1982

European Journal of Biochemistry

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Membrane vesicles from the menaquinone-deficient Bacillus subtilis aroD oxidize NADH at a low rate. NADH oxidation can be restored by the addition of slightly water-soluble menaquinone and ubiquinone analogues up to saturation levels. These saturation levels differ for the different quinone analogues tested from 95 (1,4-benzoquinone) to 5316 (5-hydroxy-1,4-naphthoquinone, juglon) nmol NADH x min-' x mg membrane protein-' N A D H oxidation in membrane vesicles from B. subtilis aroD restored with water-soluble quinone analogues supplies the energy for L-glutamate uptake. Like N A D H oxidation the initial rate of L-glutamate transport increases up to saturation levels. The highest initial rates of L-glutamate uptake are observed after restoration with quinone analogues with a relatively low standard redox potential.Functional reconstitution with natural, water-insoluble, quinones can be achieved effectively by mixing quinone-containing liposomes with membrane vesicles from B. subtilis aroD and subsequent freezing of the mixture in liquid nitrogen. The rate of N A D H oxidation increased with the amount of menaquinone incorporated in the vesicles up to saturation levels. NADH oxidation via these menaquinones also supplies the energy for L-glutamate uptake. The highest uptake rates can be obtained with menaquinone-1 and menaquinone-2.On the basis of efficiencies (mol NADH oxidized/mol L-glutamate transported) menaquinones and menaquinone analogues can be divided in two classes.To class 1 belong the menaquinone analogues, menaquinone-5 and menaquinone-8. These compounds restore NADH oxidation with low levels of energy transduction. Efficiencies are observed which are comparable with the efficiency observed in membrane vesicles from B. subtilis W23 (120-140) which contain the natural menaquinone-7.To class 2 belong menaquinone-I and menaquinone-2, which restore NADH oxidation with high levels of energy transduction. Efficiencies are observed which are in the same range as observed with phenazine methosulphate (9 -13).A model is proposed in which class 1 compounds feed in electrons from the outside from NADH to the Q-cycle of the respiratory chain. Class 2 compounds donate electrons to the respiratory chain after cytochrome c and before cytochrome a-601.Secondary transport of many solutes in membrane vesicles of Bacillus subtilis can be energized by electron transfer in the respiratory chain [l -51. The best electron donors for energization are reduced /I-nicotinamide-adenine-dinucleotide (NADH) and ascorbate/phenazine methosulphate [5 -71. A mutant has been isolated by Farrand and Taber [8,9] which is deficient in the synthesis of menaquinone, B. subtilis aroD (RB163). Membrane vesicles of this mutant oxidize NADH at a very low rate and NADH does not function as an energy source for solute transport [6,10].In order to obtain information about the structural requirements and the functional properties of menaquinones in the respiratory chain such a mutant is of particular interest. Incorporation of menaquinones in membr...

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Section: Restoration Of the Respiratory Chain Of B Subtilis Arod Witsupporting

confidence: 71%

“…In a previous publication [7] it was shown that the site of interaction of phenazine methosulphate with the respiratory chain of B. subtilis was after cytochrome ('550 and before cytochrome a601. The energy transduction via this oxidation process is about 10-fold more efficient than NADH oxidation via the class-I menaquinones.…”

Section: Discussionmentioning

confidence: 99%

Restoration of NADH Oxidation with Menaquinones and Menaquinone Analogues in Membrane Vesicles from the Menaquinone‐Deficient Bacillus subtilis aroD

Bergsma¹,

Meihuizen²,

Oeveren³

et al. 1982

European Journal of Biochemistry

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“…In co‐culture biofilms, P. aeruginosa itself may also contribute to the reduction of its own excreted 5MPCA as it does with other secreted phenazines such as PYO (Price‐Whelan et al ., 2006; 2007). We demonstrated that PMS reduction promoted uptake by C. albicans cells (Figs 6A and S7A), possibly due to an increase in its lipophilicity as the reduction of PMS makes it less soluble in aqueous solutions (Zaugg, 1964) and enhances its uptake across cell membranes (French et al ., 1973; Bisschop et al ., 1979). Our in vitro chemical studies showed that reduction of PMS is also required for spontaneous chemical conversion to red derivatives that are covalently linked to amino acids (Fig.…”

Section: Discussionmentioning

confidence: 99%

Antifungal mechanisms by which a novel Pseudomonas aeruginosa phenazine toxin kills Candida albicans in biofilms

Morales

Jacobs

Rajamani

et al. 2010

Molecular Microbiology

137

109

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SUMMARY Pseudomonas aeruginosa produces several phenazines including the recently described 5-methyl-phenazine-1-carboxylic acid (5MPCA), which exhibits a novel antibiotic activity towards pathogenic fungi such as Candida albicans. Here we characterize the unique antifungal mechanisms of 5MPCA using its analog phenazine methosulfate (PMS). Like 5MPCA, PMS induced fungal red pigmentation and killing. Mass spectrometry analyses demonstrated that PMS can be covalently modified by amino acids, a process that yields red derivatives. Furthermore, soluble proteins from C. albicans grown with either PMS or P. aeruginosa were also red and demonstrated absorbance and fluorescence spectra similar to that of PMS covalently linked to either amino acids or proteins in vitro, suggesting that 5MPCA modification by protein amine groups occurs in vivo. The red-pigmented C. albicans soluble proteins were reduced by NADH and spontaneously oxidized by oxygen, a reaction that likely generates reactive oxygen species (ROS). Additional evidence indicates that ROS generation precedes 5MPCA-induced fungal death. Reducing conditions greatly enhanced PMS fungal uptake and killing. Since 5MPCA demonstrated to be more toxic than other phenazines that are not modified, such as pyocyanin, we propose that the covalent binding of 5MPCA promotes its accumulation in target cells and contributes to its antifungal activity in mixed-species biofilms.

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“…This suggests that this may be the reason for the effects, and other potential explanations, such as changes in the MK/reduced menaquinone (MKH 2) ratio, are very unlikely. In addition, the activity of Sdh was assayed with the artificial acceptor dichlorophenol indophenol (Cl 2 Ind), which does not require the membrane anchor protein or the quinone site for reactivity [20]. In contrast to the MK-dependent activity, succinate →Cl 2 Ind activity was not inhibited by breaking the cells or adding either the uncoupler (Table 2) or valinomycin.…”

Section: Resultsmentioning

confidence: 99%

Menaquinone‐dependent succinate dehydrogenase of bacteria catalyzes reversed electron transport driven by the proton potential

Schirawski

Unden

1998

European Journal of Biochemistry

115

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Succinate dehydrogenases from bacteria and archaea using menaquinone (MK) as an electron acceptor (succinate/menaquinone oxidoreductases) contain, or are predicted to contain, two heme-B groups in the membrane-anchoring protein(s), located close to opposite sides of the membrane. All succinate/ubiquinone oxidoreductases, however, contain only one heme-B molecule. In Bacillus subtilis and other bacteria that use MK as the respiratory quinone, the succinate oxidase activity (succinate→O 2 ), and the succinate/ menaquinone oxidoreductase activity were specifically inhibited by uncoupler (CCCP, carbonyl cyanide m-chlorophenylhydrazone) or by agents dissipating the membrane potential (valinomycin). Other parts of the respiratory chains were not affected by the agents. Succinate oxidase or succinate/ubiquinone oxidoreductase from bacteria using ubiquinone as an acceptor were not inhibited. We propose that the endergonic electron transport from succinate (E°′ ϭ ϩ30 mV) to MK (E°′ Х Ϫ80 mV) in succinate/menaquinone oxidoreductase includes a reversed electron transport across the cytoplasmic membrane from the inner (negative) to the outer (positive) side via the two heme-B groups. The reversed electron transport is driven by the proton or electrical potential, which provides the driving force for MK reduction.

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Site of Interaction between Phenazine Methosulphate and the Respiratory Chain of Bacillus subtilis

Cited by 17 publications

References 21 publications

Restoration of NADH Oxidation with Menaquinones and Menaquinone Analogues in Membrane Vesicles from the Menaquinone‐Deficient Bacillus subtilis aroD

Restoration of NADH Oxidation with Menaquinones and Menaquinone Analogues in Membrane Vesicles from the Menaquinone‐Deficient Bacillus subtilis aroD

Antifungal mechanisms by which a novel Pseudomonas aeruginosa phenazine toxin kills Candida albicans in biofilms

Menaquinone‐dependent succinate dehydrogenase of bacteria catalyzes reversed electron transport driven by the proton potential

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