7RY8

S. CEREVISIAE CYP51 Y140H mutant COMPLEXED WITH Voriconazole

Summary for 7RY8

• Entry DOI: 10.2210/pdb7ry8/pdb
• Related: 4ZE3 5HS1
• Descriptor: Lanosterol 14-alpha demethylase, PROTOPORPHYRIN IX CONTAINING FE, Voriconazole, ... (6 entities in total)
• Functional Keywords: 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: 63244.42

Authors

Graham, D.O.,Wilson, R.K.,Ruma, Y.N.,Keniya, M.V.,Tyndall, J.D.,Monk, B.C. (deposition date: 2021-08-24, release date: 2021-09-01, Last modification date: 2023-10-18)

Primary citation

Graham, D.O.,Wilson, R.K.,Ruma, Y.N.,Keniya, M.V.,Tyndall, J.D.A.,Monk, B.C.
Structural Insights into the Azole Resistance of the Candida albicans Darlington Strain Using Saccharomyces cerevisiae Lanosterol 14 alpha-Demethylase as a Surrogate.
J Fungi (Basel), 7:-, 2021

PubMed Abstract: Target-based azole resistance in involves overexpression of the gene encoding lanosterol 14α-demethylase (LDM), and/or the presence of single or multiple mutations in this enzyme. Overexpression of LDM (CaLDM) Y132H I471T by the Darlington strain strongly increased resistance to the short-tailed azoles fluconazole and voriconazole, and weakly increased resistance to the longer-tailed azoles VT-1161, itraconazole and posaconazole. We have used, as surrogates, structurally aligned mutations in recombinant hexahistidine-tagged full-length LDM6×His (ScLDM6×His) to elucidate how differential susceptibility to azole drugs is conferred by LDM of the Darlington strain. The mutations Y140H and I471T were introduced, either alone or in combination, into ScLDM6×His via overexpression of the recombinant enzyme from the locus of an azole hypersensitive strain of . Phenotypes and high-resolution X-ray crystal structures were determined for the surrogate enzymes in complex with representative short-tailed (voriconazole) and long-tailed (itraconazole) triazoles. The preferential high-level resistance to short-tailed azoles conferred by the ScLDM Y140H I471T mutant required both mutations, despite the I471T mutation conferring only a slight increase in resistance. Crystal structures did not detect changes in the position/tilt of the heme co-factor of wild-type ScLDM, I471T and Y140H single mutants, or the Y140H I471T double-mutant. The mutant threonine sidechain in the Darlington strain CaLDM perturbs the environment of the neighboring C-helix, affects the electronic environment of the heme, and may, via differences in closure of the neck of the substrate entry channel, increase preferential competition between lanosterol and short-tailed azole drugs.

PubMed: 34829185
DOI: 10.3390/jof7110897

Experimental method

X-RAY DIFFRACTION (1.98 Å)