7RYX

S. CEREVISIAE CYP51 COMPLEXED WITH VT-1129

Summary for 7RYX

• Entry DOI: 10.2210/pdb7ryx/pdb
• Related: 5UL0
• Descriptor: Lanosterol 14-alpha demethylase, PROTOPORPHYRIN IX CONTAINING FE, (2R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-{5-[4-(trifluoromethoxy)phenyl]pyridin-2-yl}propan-2-ol, ... (4 entities in total)
• Functional Keywords: vt-1129, cyp51, oxidoreductase-oxidoreductase inhibitor complex, sterol biosynthesis, oxidoreductase, oxidoreductase/oxidoreductase inhibitor
• Biological source: Saccharomyces cerevisiae (strain YJM789) (Baker's yeast)
• Total number of polymer chains: 1
• Total formula weight: 62600.62

Authors

Ruma, Y.N.,Sagatova, A.,Keniya, M.V.,Tyndall, J.D.,Monk, B.C. (deposition date: 2021-08-26, release date: 2021-09-08, Last modification date: 2023-10-18)

Primary citation

Ruma, Y.N.,Keniya, M.V.,Tyndall, J.D.A.,Monk, B.C.
Characterisation of Candida parapsilosis CYP51 as a Drug Target Using Saccharomyces cerevisiae as Host.
J Fungi, 8:-, 2022

PubMed Abstract: The fungal cytochrome P450 lanosterol 14α-demethylase (CYP51) is required for the biosynthesis of fungal-specific ergosterol and is the target of azole antifungal drugs. Despite proven success as a clinical target for azole antifungals, there is an urgent need to develop next-generation antifungals that target CYP51 to overcome the resistance of pathogenic fungi to existing azole drugs, toxic adverse reactions and drug interactions due to human drug-metabolizing CYPs. is a readily transmitted opportunistic fungal pathogen that causes candidiasis in health care environments. In this study, we have characterised wild type CYP51 and its clinically significant, resistance-causing point mutation Y132F by expressing these enzymes in a host system. In some cases, the enzymes were co-expressed with their cognate NADPH-cytochrome P450 reductase (CPR). Constitutive expression of CpCYP51 Y132F conferred a 10- to 12-fold resistance to fluconazole and voriconazole, reduced to ~6-fold resistance for the tetrazoles VT-1161 and VT-1129, but did not confer resistance to the long-tailed triazoles. Susceptibilities were unchanged in the case of CpCPR co-expression. Type II binding spectra showed tight triazole and tetrazole binding by affinity-purified recombinant CpCYP51. We report the X-ray crystal structure of ScCYP51 in complex with VT-1129 obtained at a resolution of 2.1 Å. Structural analysis of azole-enzyme interactions and functional studies of recombinant CYP51 from have improved understanding of their susceptibility to azole drugs and will help advance structure-directed antifungal discovery.

PubMed: 35050009
DOI: 10.3390/jof8010069

Experimental method

X-RAY DIFFRACTION (2.1 Å)