Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1990 Mar 1;266(2):475–480. doi: 10.1042/bj2660475

Interaction of azole antifungal antibiotics with cytochrome P-450-dependent 14 alpha-sterol demethylase purified from Candida albicans.

C A Hitchcock 1, K Dickinson 1, S B Brown 1, E G Evans 1, D J Adams 1
PMCID: PMC1131156  PMID: 2180400

Abstract

The interaction of azole antifungal antibiotics with purified Candida albicans cytochrome P-450-dependent 14 alpha-sterol demethylase (P-450DM) was measured spectrophotometrically and by inhibition of enzyme activity. Ketoconazole and ICI 153066 (a triazole derivative) formed low-spin complexes with the ferric cytochrome and induced type II difference spectra. These spectra are indicative of an interaction between the azole moiety and the sixth co-ordination position of P-450DM haem. Both azoles inhibited the binding of CO to the sodium dithionite-reduced ferrous cytochrome, and inhibited reconstituted P-450DM activity by binding to the cytochrome with a one-to-one stoichiometry. Similarly, total inhibition of enzyme activity occurred when equimolar amounts of clotrimazole, miconazole or fluconazole were added to reconstituted P-450DM. These results correlated with the inhibition of P-450DM in broken cell preparations, confirming that all five azoles are potent inhibitors of ergosterol biosynthesis in C. albicans.

Full text

PDF
475

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aoyama Y., Yoshida Y., Hata S., Nishino T., Katsuki H. Buthiobate: a potent inhibitor for yeast cytochrome P-450 catalyzing 14 alpha-demethylation of lanosterol. Biochem Biophys Res Commun. 1983 Sep 15;115(2):642–647. doi: 10.1016/s0006-291x(83)80192-2. [DOI] [PubMed] [Google Scholar]
  2. Aoyama Y., Yoshida Y., Sato R. Yeast cytochrome P-450 catalyzing lanosterol 14 alpha-demethylation. II. Lanosterol metabolism by purified P-450(14)DM and by intact microsomes. J Biol Chem. 1984 Feb 10;259(3):1661–1666. [PubMed] [Google Scholar]
  3. Aoyama Y., Yoshida Y., Sonoda Y., Sato Y. Metabolism of 32-hydroxy-24,25-dihydrolanosterol by purified cytochrome P-45014DM from yeast. Evidence for contribution of the cytochrome to whole process of lanosterol 14 alpha-demethylation. J Biol Chem. 1987 Jan 25;262(3):1239–1243. [PubMed] [Google Scholar]
  4. Ardies C. M., Lasker J. M., Bloswick B. P., Lieber C. S. Purification of NADPH:cytochrome c (cytochrome P-450) reductase from hamster liver microsomes by detergent extraction and affinity chromatography. Anal Biochem. 1987 Apr;162(1):39–46. doi: 10.1016/0003-2697(87)90008-x. [DOI] [PubMed] [Google Scholar]
  5. Barrett-Bee K. J., Lane A. C., Turner R. W. The mode of antifungal action of tolnaftate. J Med Vet Mycol. 1986 Apr;24(2):155–160. doi: 10.1080/02681218680000221. [DOI] [PubMed] [Google Scholar]
  6. Fromtling R. A. Overview of medically important antifungal azole derivatives. Clin Microbiol Rev. 1988 Apr;1(2):187–217. doi: 10.1128/cmr.1.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hitchcock C. A., Barrett-Bee K. J., Russell N. J. Inhibition of 14 alpha-sterol demethylase activity in Candida albicans Darlington does not correlate with resistance to azole. J Med Vet Mycol. 1987 Oct;25(5):329–333. [PubMed] [Google Scholar]
  8. Hitchcock C. A., Brown S. B., Evans E. G., Adams D. J. Cytochrome P-450-dependent 14 alpha-demethylation of lanosterol in Candida albicans. Biochem J. 1989 Jun 1;260(2):549–556. doi: 10.1042/bj2600549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hitchcock C. A., Dickinson K., Brown S. B., Evans E. G., Adams D. J. Purification and properties of cytochrome P-450-dependent 14 alpha-sterol demethylase from Candida albicans. Biochem J. 1989 Oct 15;263(2):573–579. doi: 10.1042/bj2630573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kalb V. F., Woods C. W., Turi T. G., Dey C. R., Sutter T. R., Loper J. C. Primary structure of the P450 lanosterol demethylase gene from Saccharomyces cerevisiae. DNA. 1987 Dec;6(6):529–537. doi: 10.1089/dna.1987.6.529. [DOI] [PubMed] [Google Scholar]
  11. Lai M. H., Kirsch D. R. Nucleotide sequence of cytochrome P450 L1A1 (lanosterol 14 alpha-demethylase) from Candida albicans. Nucleic Acids Res. 1989 Jan 25;17(2):804–804. doi: 10.1093/nar/17.2.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964 Jul;239:2370–2378. [PubMed] [Google Scholar]
  13. Pye G. W., Marriott M. S. Inhibition of sterol C14 demethylation by imidazole-containing antifungals. Sabouraudia. 1982 Dec;20(4):325–329. doi: 10.1080/00362178285380461. [DOI] [PubMed] [Google Scholar]
  14. Trzaskos J., Kawata S., Gaylor J. L. Microsomal enzymes of cholesterol biosynthesis. Purification of lanosterol 14 alpha-methyl demethylase cytochrome P-450 from hepatic microsomes. J Biol Chem. 1986 Nov 5;261(31):14651–14657. [PubMed] [Google Scholar]
  15. Vanden Bossche H. Biochemical targets for antifungal azole derivatives: hypothesis on the mode of action. Curr Top Med Mycol. 1985;1:313–351. doi: 10.1007/978-1-4613-9547-8_12. [DOI] [PubMed] [Google Scholar]
  16. White R. E., Coon M. J. Heme ligand replacement reactions of cytochrome P-450. Characterization of the bonding atom of the axial ligand trans to thiolate as oxygen. J Biol Chem. 1982 Mar 25;257(6):3073–3083. [PubMed] [Google Scholar]
  17. Yeagle P. L., Martin R. B., Lala A. K., Lin H. K., Bloch K. Differential effects of cholesterol and lanosterol on artificial membranes. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4924–4926. doi: 10.1073/pnas.74.11.4924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yoshida Y., Aoyama Y. Interaction of azole antifungal agents with cytochrome P-45014DM purified from Saccharomyces cerevisiae microsomes. Biochem Pharmacol. 1987 Jan 15;36(2):229–235. doi: 10.1016/0006-2952(87)90694-0. [DOI] [PubMed] [Google Scholar]
  19. Yoshida Y., Aoyama Y., Takano H., Kato T. Stereo-selective interaction of enantiomers of diniconazole, a fungicide, with purified P-450/14DM from yeast. Biochem Biophys Res Commun. 1986 May 29;137(1):513–519. doi: 10.1016/0006-291x(86)91240-4. [DOI] [PubMed] [Google Scholar]
  20. Yoshida Y., Aoyama Y. Yeast cytochrome P-450 catalyzing lanosterol 14 alpha-demethylation. I. Purification and spectral properties. J Biol Chem. 1984 Feb 10;259(3):1655–1660. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES