| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
Papers In Press, published online ahead of print August 16, 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Department of Biochemistry, Vanderbilt University, Nashville, TN 37232-0146
Corresponding Author: aouatef.bellamine{at}vanderbilt.edu
14a-Demethylase (CYP51) is a key enzyme in all sterol biosynthetic pathways (animals, fungi, plants, protists and some bacteria) catalyzing the removal of the C-14 methyl group following cyclization of squalene. Based on mutations found in CYP51 genes from Candida albicans azole-resistant isolates obtained after fluconazole treatment of fungal infections, and using site-direct mutagenesis, we have found that fluconazole binding and substrate metabolism vary among three different CYP51 isoforms, human, fungal and mycobacterial. In C. albicans, the Y132H mutant from isolates shows no effect on fluconazole binding whereas the F145L mutant results in 5-fold increase in its IC50 for fluconazole, suggesting that F145 (conserved only in fungal 14a-demethylases) interacts with this azole. In C. albicans, F145L accounts, in part, for the difference in fluconazole sensitivity reported between mammals and fungi providing a basis for treatment of fungal infections. While the C. albicans Y132H and human Y145H CYP51 mutants demonstrate essentially no effect on substrate metabolism, the Mycobacterium tuberculosis F89H CYP51 mutant loses both its substrate binding and metabolism. Since these three residues align in the three isoforms, the results indicate that their active sites contain important structural differences, and further emphasizes that fluconazole and substrate binding are uncoupled properties
Revised on August 9, 2004
Accepted on August 11, 2004
Fluconazole binding and sterol demethylation in three CYP51 isoforms indicate differences in active site topology
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. E. Parker, M. Merkamm, N. J. Manning, D. Pompon, S. L. Kelly, and D. E. Kelly Differential Azole Antifungal Efficacies Contrasted Using a Saccharomyces cerevisiae Strain Humanized for Sterol 14{alpha}-Demethylase at the Homologous Locus Antimicrob. Agents Chemother., October 1, 2008; 52(10): 3597 - 3603. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. N. Eddine, J. P. von Kries, M. V. Podust, T. Warrier, S. H. E. Kaufmann, and L. M. Podust X-ray Structure of 4,4'-Dihydroxybenzophenone Mimicking Sterol Substrate in the Active Site of Sterol 14{alpha}-Demethylase (CYP51) J. Biol. Chem., May 30, 2008; 283(22): 15152 - 15159. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kudo, M. Ohi, Y. Aoyama, Y. Nitahara, S.-K. Chung, and Y. Yoshida 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 J. Biochem., May 1, 2005; 137(5): 625 - 632. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Matsuura, S. Yoshioka, T. Tosha, H. Hori, K. Ishimori, T. Kitagawa, I. Morishima, N. Kagawa, and M. R. Waterman Structural Diversities of Active Site in Clinical Azole-bound Forms between Sterol 14{alpha}-Demethylases (CYP51s) from Human and Mycobacterium tuberculosis J. Biol. Chem., March 11, 2005; 280(10): 9088 - 9096. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Journal of Biological Chemistry |
| Molecular and Cellular Proteomics | ASBMB Today |
