Disruption of ergosterol biosynthesis, growth, and the morphological transition in Candida albicans by sterol methyltransferase inhibitors containing sulfur at C-25 in the sterol side chain.
The sterol substrate analog 25-thialanosterol and its corresponding sulfonium salt were evaluated for their ability to serve as antifungal agents and to inhibit sterol methyltransferase (SMT) activity in Candida albicans. Both compounds inhibited cell proliferation, were fungistatic, interrupted the yeast-like-form to germ-tube-form transition, and resulted in the accumulation of zymosterol and related delta24-sterols concurrent with a decrease in ergosterol, as was expected for the specific inhibition of SMT activity. Feedback on sterol synthesis was evidenced by elevated levels of cellular sterols in treated vs. control cultures. However, neither farnesol nor squalene accumulated in significant amounts in treated cultures, suggesting that carbon flux is channeled from the isoprenoid pathway to the sterol pathway with minor interruption or redirection until blockage at the C-methylation step. Activity assays using solubilized C. albicans SMT confirmed the inhibitors impair SMT action. Kinetic analysis indicated that 25-thialanosterol inhibited SMT with the properties of a time-dependent mechanism-based inactivator Ki of 5 microM and apparent kinact of 0.013 min(-1), whereas the corresponding sulfonium salt was a reversible-type transition state analog exhibiting a Ki of 20 nM. The results are interpreted to imply changes in ergosterol homeostasis as influenced by SMT activity can control growth and the morphological transition in C. albicans, possibly affecting disease development.
Kanagasabai
Ragu
R
Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas 79409, USA.
Zhou
Wenxu
W
Liu
Jialin
J
Nguyen
Thi Thuy Minh
TT
Veeramachaneni
Phani
P
Nes
W David
WD
D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D002176:Candida albicans / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D049109:Cell Proliferation; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D004875:Ergosterol / Q000096:biosynthesis* / Q000737:chemistry; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008745:Methylation; D008780:Methyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D015394:Molecular Structure; D013261:Sterols / Q000737:chemistry / Q000378:metabolism*; D013455:Sulfur / Q000737:chemistry
D000935:Antifungal Agents; D004791:Enzyme Inhibitors; D013261:Sterols; D013455:Sulfur; D008780:Methyltransferases; D004875:Ergosterol