Long-chain alkyl ester of AMP acts as an antagonist of glucose-induced signal transduction that mediates activation of plasma membrane proton pump in Saccharomyces cerevisiae

Abstract

One of the long-chain alkyl esters of AMP, adenosine 5'-hexadecylphosphate (AMPC16), exhibited a cytotoxic growth inhibitory effect on cells of various yeast strains. The growth inhibitory effect of AMPC16 on Saccharomyces cerevisiae cells was observed only in medium containing Mg 2M , which accelerated cellular uptake of the nucleotide analogue. In the presence of Mg 2M , AMPC16 completely inhibited glucose-induced extracellular acidification by the intact cells and also interfered with activation of the plasma membrane ATPase, but did not directly inhibit the ATPase activity itself. AMPC16 treatment prevented cells from increasing their intracellular sn-1,2-diacylglycerol (DAG) level in response to glucose, whereas the inhibition of proton extrusion by the cells could be largely reversed by the coaddition of a membrane-permeable DAG analogue. The DAG analogue, a physiological activator of protein kinase C (PKC), was not protective against the inhibition of glucose-induced proton extrusion by staurosporine, which is capable of directly interfering with the action of PKC. These results implied that AMPC16 caused a Mg 2M -dependent cytotoxic effect on Sac. cerevisiae cells by interfering with a phosphatidylinositol type of signal that mediates activation of the plasma membrane proton pump.

The Synthetic Amphipathic Peptidomimetic LTX109 Is a Potent Fungicide That Disturbs Plasma Membrane Integrity in a Sphingolipid Dependent Manner

“…Glucose-induced acidification was measured as previously described [32] with modifications. Exponentially growing S. cerevisiae cells were washed and resuspended in sterile water to a final concentration of 10 8 cells/ml.…”

Synergistic Fungicidal Activities of Amphotericin B and N-Methyl-N″-dodecylguanidine: A Constituent of Polyol Macrolide Antibiotic Niphimycin

“…MICs of N-methyl-NЉ-alkylguanidines, fatty acids, and alkylamines were determined for S. cerevisiae X 2180-1A (a) in the presence or absence of AmB by the serial 2-fold broth dilution method as follows [6]. After diluting an overnight culture with YPD medium (yeast extract 1%, peptone 2%, and glucose 2%) to 10 6 cells/ml, the cell suspension was incubated in a freshly prepared medium in a 96-well microplate at 30°C for 48 hours.…”

Visualization analysis of the vacuole-targeting fungicidal activity of amphotericin B against the parent strain and an ergosterol-less mutant of Saccharomyces cerevisiae

“…Minimum growth inhibitory concentrations (MICs) of AmB, AmBDs and MC12 were determined by the twofold broth dilution method, applying the chequerboard technique (Davis et al, 1994;Tanaka et al, 2000). Cells were grown overnight in YPD medium containing 1 % yeast extract (Difco Laboratories), 2 % bacto-peptone (Difco Laboratories), and 2 % D-glucose, with vigorous shaking at 30 uC.…”