The inhibitory effect of the artificial electron donor system, phenazine methosulfate‐ascorbate, on bacterial transport mechanisms

Abstract

The artificial electron donor system, phenazine methosulfate (PMS)-ascorbate, inhibited active transort of solutes in Pseudomonas aeruginosa irrespective of whether the active transport systems were shock sensitive or shock resistant. N,N,N',N'-tetramethylphenylenediamine could be substituted for PMS but a higher concentration was required. PMS-ascorbate also inhibited active transport in several other bacterial species with the exception of Escherichia coli and of a nonpigmented strain of Serratia marcescens. PMS-ascorbate previously has been shown to energize active transport in isolated membrane vesicles, even those prepared from the same bacterial species in whose intact cells active transport was inhibited. The apparent Km of glucose active transport in untreated cells of P. aeruginosa was 40 micron while the Km of glucose transport in cells incubated with PMS-ascorbate was 25 mM, and PMS-ascorbate had no effect on efflux of accumulated glucose. These results strongly suggested that facilitated diffusion resulted upon exposure of the cells to PMS- ascorbate. Thus, PMS-ascorbate appeared to have an uncoupler-like effect on cells of P. aeruginosa. The experimental data also pointed out that there are fundamental differences between the response of intact cells and membrane vesicles to exogenous electron donors.

Stimulating Effect of an Artificial Electron Mediator, Phenazine Methosulfate, on Rb<SUP>+</SUP> Transport in HeLa Cells

“…Since Singer and Kearney introduced PMS as an intermediate electron carrier to link succinate dehydrogenase to oxygen (23), this phenazine dye has been used as a practical electron mediator to couple various dehydrogenases to the physiological or artificial electron donors or acceptors. PMS has been reported to affect the membrane transport of solutes and the metabolism of mammalian (2,3,8,11,25), and bacterial cells (1,6,7,12,21). With respect to monovalent cation transport, PMSascorbate stimulates the valinomycin-induced concentrative Rb+ uptake into vesicles of the bacterial membrane (10,13,14).…”