Effect of miconazole on the structure and function of plasma membrane of Candida albicans.
The primary mode of action of azoles is the inhibition of cytochrome P-450 dependent 14 alpha-demethylase, a key enzyme in ergosterol biosynthesis in fungi. Our results demonstrated that Candida albicans cells grown in the presence of 10 micrograms ml-1 of miconazole (miconazole-grown), do not possess ergosterol in their plasma membranes and this ergosterol depletion leads to a drastic change in membrane fluidity as shown by fluorescence polarization measurements and unsaturation index. There was an increase in membrane order in miconazole-grown cells and a reduced rate of uptake of amino acids. We also checked for membrane permeability changes in normal mid-log phase cells (normal-grown) in short incubations (10 min) with 10 micrograms ml-1 miconazole (miconazole-incubated). Interestingly, the amino acid uptake rates except that of Gly were not affected significantly in these cells. The results suggest that in the miconazole-incubated cells, the drug is not able to alter the level of ergosterol or inhibit ergosterol biosynthesis during 10 min incubation and therefore the interaction of the drug neither leads to significant disorganization of membrane components, nor affects permease activity, whereas in the miconazole-grown cells there is ergosterol depletion leading to accumulation of biosynthetic intermediates, resulting in membrane rearrangement thereby causing a major fluidity change. This fluidity change may explain the drastic reduction of amino acid transport in miconazole-grown cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Ansari
S
S
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
Prasad
R
R
D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D002176:Candida albicans / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D004875:Ergosterol / Q000032:analysis; D005227:Fatty Acids / Q000032:analysis; D005454:Fluorescence Polarization; D008560:Membrane Fluidity / Q000187:drug effects / Q000502:physiology; D008825:Miconazole / Q000494:pharmacology*; D010743:Phospholipids / Q000032:analysis; D013104:Spheroplasts / Q000737:chemistry / Q000187:drug effects; D013261:Sterols / Q000032:analysis
D000596:Amino Acids; D005227:Fatty Acids; D010743:Phospholipids; D013261:Sterols; D008825:Miconazole; D004875:Ergosterol