Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites

Microbiology (Reading). 2017 Mar;163(3):364-372. doi: 10.1099/mic.0.000429. Epub 2017 Mar 29.

Abstract

Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 µM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3 % of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Anthracenes / pharmacology*
  • Antifungal Agents / pharmacology*
  • Candida albicans / drug effects
  • Candida albicans / growth & development
  • Cell Line, Tumor
  • Cell Respiration / drug effects*
  • Cucumis sativus / drug effects
  • Energy Metabolism / drug effects*
  • Humans
  • Magnetic Resonance Spectroscopy
  • Microbial Sensitivity Tests
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Mycelium / drug effects
  • Mycelium / growth & development
  • Mycelium / metabolism
  • Niflumic Acid / pharmacology*
  • Patch-Clamp Techniques
  • Phycomyces / drug effects
  • Phycomyces / growth & development*
  • Phycomyces / metabolism
  • Polyphosphates / metabolism
  • Voltage-Dependent Anion Channels / antagonists & inhibitors

Substances

  • Anthracenes
  • Antifungal Agents
  • Polyphosphates
  • Voltage-Dependent Anion Channels
  • Niflumic Acid
  • 9-anthroic acid
  • Adenosine Triphosphate