Three-dimensional models of wild-type and mutated forms of cytochrome P450 14alpha-sterol demethylases from Aspergillus fumigatus and Candida albicans provide insights into posaconazole binding.
The cytochrome P450 sterol 14alpha-demethylase enzyme (CYP51) is the target of azole antifungals. Azoles block ergosterol synthesis, and thereby fungal growth, by binding in the active-site cavity of the enzyme and ligating the iron atom of the heme cofactor through a nitrogen atom of the azole. Mutations in and around the CYP51 active site have resulted in azole resistance. In this work, homology models of the CYP51 enzymes from Aspergillus fumigatus and Candida albicans were constructed based on the X-ray crystal structure of CYP51 from Mycobacterium tuberculosis. Using these models, binding modes for voriconazole (VOR), fluconazole (FLZ), itraconazole (ITZ), and posaconazole (POS) were predicted from docking calculations. Previous work had demonstrated that mutations in the vicinity of the heme cofactor had a greater impact on the binding of FLZ and VOR than on the binding of POS and ITZ. Our modeling data suggest that the long side chains of POS and ITZ occupy a specific channel within CYP51 and that this additional interaction, which is not available to VOR and FLZ, serves to stabilize the binding of these azoles to the mutated CYP51 proteins. The model also predicts that mutations that were previously shown to specifically impact POS susceptibility in A. fumigatus and C. albicans act by interfering with the binding of the long side chain.
Xiao
Li
L
Department of Structural Chemistry, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA. li.xiao@spcorp.com
Madison
Vincent
V
Chau
Andrew S
AS
Loebenberg
David
D
Palermo
Robert E
RE
McNicholas
Paul M
PM
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000378:metabolism* / Q000494:pharmacology; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D001393:Azoles / Q000494:pharmacology; D001665:Binding Sites; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics*; D025141:Drug Resistance, Fungal; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics*; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics / Q000502:physiology; D011485:Protein Binding; D014230:Triazoles / Q000378:metabolism*
D000935:Antifungal Agents; D001393:Azoles; D005656:Fungal Proteins; D008024:Ligands; D008565:Membrane Proteins; D014230:Triazoles; C101425:posaconazole; D003577:Cytochrome P-450 Enzyme System; C465527:cytochrome P-450 CYP51B, Aspergillus