In Silico Structural Modeling and Analysis of Interactions of Tremellomycetes Cytochrome P450 Monooxygenases CYP51s with Substrates and Azoles

Abstract

Cytochrome P450 monooxygenase CYP51 (sterol 14α-demethylase) is a well-known target of the azole drug fluconazole for treating cryptococcosis, a life-threatening fungal infection in immune-compromised patients in poor countries. Studies indicate that mutations in CYP51 confer fluconazole resistance on cryptococcal species. Despite the importance of CYP51 in these species, few studies on the structural analysis of CYP51 and its interactions with different azole drugs have been reported. We therefore performed in silico structural analysis of 11 CYP51s from cryptococcal species and other Tremellomycetes. Interactions of 11 CYP51s with nine ligands (three substrates and six azoles) performed by Rosetta docking using 10,000 combinations for each of the CYP51-ligand complex (11 CYP51s × 9 ligands = 99 complexes) and hierarchical agglomerative clustering were used for selecting the complexes. A web application for visualization of CYP51s’ interactions with ligands was developed (http://bioshell.pl/azoledocking/). The study results indicated that Tremellomycetes CYP51s have a high preference for itraconazole, corroborating the in vitro effectiveness of itraconazole compared to fluconazole. Amino acids interacting with different ligands were found to be conserved across CYP51s, indicating that the procedure employed in this study is accurate and can be automated for studying P450-ligand interactions to cater for the growing number of P450s.

Mulberrin inhibitsBotrytis cinereafor strawberry storage by interfering with the bioactivity of 14α-demethylase (CYP51)

“…After the medium was solidified, B. cinerea with a diameter of 0.0141 m (cut from the edge of the colony) was inoculated in the dish center and cultured at 28 °C until the colonies in the mock treatment grew to the dish edge. Phosphate buffered saline (PBS buffer, 256 μg mL −1 ) and fluconazole [26][27][28] (128 μg mL −1 ) were used to replace the four natural compound solutions with mock treatment and positive control, respectively. Each experiment was repeated five times.…”

Lack of Association between Fluconazole Susceptibility and ERG11 Nucleotide Polymorphisms in Cryptococcus neoformans Clinical Isolates from Uganda

“…Heme binding sites are highlighted in purple [ 20 ]. Substrate recognition sites are highlighted in green [ 45 ].…”