5ESK

Saccharomyces cerevisiae CYP51 (Lanosterol 14-alpha demethylase) G464S mutant complexed with itraconazole

Summary for 5ESK

• Entry DOI: 10.2210/pdb5esk/pdb
• Related: 5ESE 5ESF 5ESG 5ESH 5ESI 5ESJ 5ESL 5ESM 5ESN
• Descriptor: Lanosterol 14-alpha demethylase, 2-[(2R)-butan-2-yl]-4-{4-[4-(4-{[(2R,4S)-2-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolan-4-yl]methoxy}phenyl)piperazin-1-yl]phenyl}-2,4-dihydro-3H-1,2,4-triazol-3-one, PROTOPORPHYRIN IX CONTAINING FE, ... (4 entities in total)
• Functional Keywords: cyp51, oxidoreductase-oxidoreductase inhibitor complex, oxidoreductase/oxidoreductase inhibitor
• Biological source: Saccharomyces cerevisiae (strain YJM789) (Baker's yeast)
• Total number of polymer chains: 1
• Total formula weight: 63237.36

Authors

Sagatova, A.,Keniya, M.V.,Wilson, R.K.,Sabherwal, M.,Tyndall, J.D.A.,Monk, B.C. (deposition date: 2015-11-16, release date: 2016-11-23, Last modification date: 2023-09-27)

Primary citation

Sagatova, A.A.,Keniya, M.V.,Tyndall, J.D.A.,Monk, B.C.
Impact of Homologous Resistance Mutations from Pathogenic Yeast on Saccharomyces cerevisiae Lanosterol 14 alpha-Demethylase.
Antimicrob.Agents Chemother., 62:-, 2018

PubMed Abstract: Fungal infections frequently affect immunodeficient individuals and are estimated to kill 1.35 million people per annum. Azole antifungals target the membrane-bound cytochrome P450 monooxygenase lanosterol 14α-demethylase (CYP51; Erg11p). Mutations in CYP51 can render the widely used triazole drugs less effective. The CYP51 mutation G464S and the double mutation Y132F G464S (Y140F and G464S by numbering) as well as the CYP51A G54E/R/W mutations of (G73E/R/W by numbering) have been reproduced in a recombinant C-terminal hexahistidine-tagged version of CYP51 (ScErg11p6×His). Phenotypes and X-ray crystal structures were determined for the mutant enzymes. Liquid microdilution assays showed that the G464S mutation in ScErg11p6×His conferred no difference in the susceptibility of yeast to triazole drugs but in combination with the Y140F mutation gave a 4-fold reduction in susceptibility to the short-tailed triazole fluconazole. The ScErg11p6×His Y140F G464S mutant was unstable during purification and was not crystallized. The ScErg11p6×His G73E/R/W mutations conferred increased susceptibly to all triazoles tested in liquid microdilution assays. High-resolution X-ray crystal structures reveal two different conformations of the ligand itraconazole, including a previously unseen conformation, as well as interactions between the tail of this triazole and the E/W73 residue. This study shows that CYP51 adequately represents some but not all mutations in CYP51s of pathogenic fungi. Insight into the molecular mechanisms of resistance mutations in CYP51 will assist the development of inhibitors that will overcome antifungal resistance.

PubMed: 29263059
DOI: 10.1128/AAC.02242-17

Experimental method

X-RAY DIFFRACTION (2.24 Å)