Enhancement of the Fungicidal Activity of Amphotericin B by Allicin: Effects on Intracellular Ergosterol Trafficking

Abstract

In Saccharomyces cerevisiae, ergosterol was visible in the plasma membrane of untreated cells and was enriched in the vacuole membrane in response to amphotericin B (AmB) treatment at a non-lethal concentration. The simultaneous addition of allicin was inhibitory to AmB-induced ergosterol enrichment in the vacuole membrane, resulting in increased sensitivity of the organelles to the disruptive action of AmB. Allicin was also inhibitory to ergosterol enrichment in the vacuole membrane that was achieved by its external addition to cells. The combined fungicidal activity of AmB and allicin was suppressed together with suppression of vacuole membrane damage in cells where ergosterol had been fully enriched in the vacuole membrane. AmB caused direct disruptive damage of the isolated vacuoles, but the antibiotic was apparently less effective in disrupting the organelles that were isolated after ergosterol enrichment. These findings suggest that allicin enhances AmB-induced vacuole membrane damage by inhibiting ergosterol trafficking from the plasma membrane to the vacuole membrane.

Key words

allicin - amphotericin B - ergosterol trafficking - vacuole morphology - Saccharomyces cerevisiae

References

• 1 Brajtburg J, Powderly W G, Kobayashi G S, Medoff G. Amphotericin B: current understanding of mechanisms of action.  Antimicrob Agents Chemother. 1999;  34 183-8
• 2 Chen W C, Chou D L, Feingold D S. Dissociation between ion permeability and the lethal action of polyene antibiotics on Candida albicans. .  Antimicrob Agents Chemother. 1978;  13 914-7
• 3 Alonso M A, Vázquez D, Carrasco L. Compounds affecting membranes that inhibit protein synthesis in yeast.  Antimicrob Agents Chemother. 1979;  16 750-6
• 4 Bhuiyan M S, Ito Y, Nakamura A, Tanaka N, Fujita K, Fukui H. et al . Nystatin effects on vacuolar function in Saccharomyces cerevisiae. .  Biosci Biotechnol Biochem. 1999;  63 1075-82
• 5 Ogita A, Fujita K, Taniguchi M, Tanaka T. Enhancement of the fungicidal activity of amphotericin B by allicin, an allyl-sulfur compound from garlic, against the yeast Saccharomyces cerevisiae as a model system.  Planta Med. 2006;  72 1247-50
• 6 Ankri S, Mirelman D. Antimicrobial properties of allicin from garlic.  Microbes Infect. 1999;  1 125-9
• 7 Oommen S, Anto R J, Srinivas G, Karunagaran D. Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells.  Eur J Pharmacol. 2004;  485 97-103
• 8 Ogita A, Hirooka K, Yamamoto Y, Tsutsui N, Fujita K, Taniguchi M. et al . Synergistic fungicidal activity of Cu²⁺ and allicin, an allyl sulfur compound from garlic, and its relation to the role of alkyl hydroperoxide reductase 1 as a cell surface defense in Saccharomyces cerevisiae. .  Toxicology. 2005;  215 205-13
• 9 Ogita A, Fujita K, Taniguchi M, Tanaka T. Dependence of synergistic fungicidal activity of Cu²⁺ and allicin, an allyl sulfur compound from garlic, on selective accumulation of the ion in the plasma membrane fraction via allicin-mediated phospholipid peroxidation.  Planta Med. 2006;  72 875-80
• 10 Ogita A, Nagao Y, Fujita K, Tanaka T. Amplification of vacuole-targeting fungicidal activity of antibacterial antibiotic polymyxin B by allicin, an allyl sulfur compound from garlic.  J Antibiot. 2007;  60 511-8
• 11 Ogita A, Matsumoto K, Fujita K, Usuki Y, Hatanaka Y, Tanaka T. Synergistic fungicidal activities of amphotericin B and N-methyl-N″-dodecylguanidine: a constituent of polyol macrolide antibiotic niphimycin.  J Antibiot. 2007;  60 27-35
• 12 Beh C T, Rine J. A role for yeast oxysterol-binding protein homologs in endocytosis and in the maintenance of intracellular sterol-lipid distribution.  J Cell Sci. 2004;  15 2983-96
• 13 Conradt B, Shaw J, Vida T, Emr S, Wickner W. In vitro reactions of vacuole inheritance in Saccharomyces cerevisiae. .  J Cell Biol. 1992;  119 469-79
• 14 Ohsumi Y, Anraku Y. Active transport of basic amino acids driven by a proton motive force in vacuolar membrane vesicles of Saccharomyces cerevisiae. .  J Biol Chem. 1981;  256 2079-82
• 15 Tabuchi M, Iwaihara O, Ohtani Y, Ohuchi N, Sakurai J, Morita T. et al . Vacuolar protein sorting in fission yeast: cloning, biosynthesis, transport, and processing of carboxypeptidase Y from Schizosaccharomyces pombe. .  J Bacteriol. 1997;  179 4179-89
• 16 Kato M, Wickner W. Ergosterol is required for the Sec18/ATP-dependent priming step of homotypic vacuole fusion.  EMBO J. 2001;  20 4035-40
• 17 Tedrick K, Trischuk T, Lehner R, Eitzen G. Enhanced membrane fusion in sterol-enriched vacuoles bypasses the Vrp1 p requirement.  Mol Biol Cell. 2004;  15 4609-21
• 18 Munn A L, Heese-Peck A, Stevenson B J, Pichler H, Riezman H. Specific sterols required for the internalization step of endocytosis in yeast.  Mol Biol Cell. 1999;  10 3943-57
• 19 Heese-Peck A, Pichler H, Zanolari B, Watanabe R, Daum G, Riezman H. Multiple functions of sterols in yeast endocytosis.  Mol Biol Cell. 2002;  13 2664-80
• 20 Hitchcock C A, Barrett-Bee K J, Russell N J. The lipid composition and permeability to azole of an azole- and polyene-resistant mutant of Candida albicans. .  J Med Vet Mycol. 1987;  25 29-37
• 21 Subden R E, Safe L, Morris D C, Brown R G, Safe S. Eburicol, lichosterol, ergosterol and obtusifoliol from polyene antibiotic-resistant mutants of Candida albicans. .  Can J Microbiol. 1977;  23 751-4
• 22 Athar M A, Winner H L. The development of resistance by Candida albicans to polyene antibiotics in vitro. .  J Med Microbiol. 1971;  4 505-17
• 23 Hamilton-Miller J MT. Sterols from polyene-resistant mutants of Candida albicans. .  J Gen Microbiol. 1972;  73 201-3
• 24 Pierce A M, Pierce HD J r., Unrau A M, Oehlschlager A C. Lipid composition and polyene antibiotic resistance of Candida albicans. .  Can J Biochem. 1978;  56 135-42
• 25 Boller T, Dürr M, Wiemken A. Characterization of a specific transport system for arginine in isolated yeast vacuoles.  Eur J Biochem. 1975;  54 81-91