Effects of Y132H and F145L substitutions on the activity, azole resistance and spectral properties of Candida albicans sterol 14-demethylase P450 (CYP51): a live example showing the selection of altered P450 through interaction with environmental compounds.
Three variants of Candida albicans CYP51 (sterol 14-demethylase P450) having Y132H and/or F145L substitutions were purified and characterized to reveal the effects of these amino acid substitutions on the enzymatic properties and azole resistance of the enzyme. Y132H and F145L substitutions modified the spectral properties of the enzyme, suggesting that they caused some structural change modifying the heme environments of CYP51. Y132H and F145L substitutions increased the resistance of the enzyme to azole compounds but considerably decreased the catalytic activity. This fact represents a trade-off between acquisition of azole resistance and maintenance of high activity in the CYP51 having Y132H and F145L substitutions. A fluconazole-resistant C. albicans strain DUMC136 isolated from patients receiving long-term azole treatment was a homozygote of the altered CYP51 having Y132H and F145L substitutions. However, neither of these substitutions was found in CYP51 of wild-type C. albicans so far studied. These facts suggest that the azole-resistant variant having Y132H and/or F145L substitutions might be selected only under azole-rich environments because of its azole resistance and impaired catalytic activity. This may be a live example showing one of the important processes of P450 diversification, the selection of altered P450 through the interaction with environmental compounds.
Kudo
Makiko
M
School of Pharmaceutical Sciences and Institute for Bioscience, Mukogawa Women's University, Nishinomiya, Hyogo 663-8179.
Ohi
Miwa
M
Aoyama
Yuri
Y
Nitahara
Yuko
Y
Chung
Sung-Kee
SK
Yoshida
Yuzo
Y
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects* / Q000235:genetics*; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004227:Dithionite / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015725:Fluconazole / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics*; D006418:Heme / Q000737:chemistry; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism; D013057:Spectrum Analysis; D058886:Sterol 14-Demethylase
D000935:Antifungal Agents; D001393:Azoles; D065607:Cytochrome P-450 Enzyme Inhibitors; D005656:Fungal Proteins; C465525:cytochrome P-450 CYP51, Candida albicans; D004227:Dithionite; D006418:Heme; D015725:Fluconazole; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D058886:Sterol 14-Demethylase