5EAB

Saccharomyces cerevisiae CYP51 complexed with the plant pathogen inhibitor S-tebuconazole

5EAB の概要

• エントリーDOI: 10.2210/pdb5eab/pdb
• 関連するPDBエントリー: 4WMZ 5EAC 5EAD 5EAE 5EAF 5EAG 5EAH
• 分子名称: Lanosterol 14-alpha demethylase, PROTOPORPHYRIN IX CONTAINING FE, (3~{S})-1-(4-chlorophenyl)-4,4-dimethyl-3-(1,2,4-triazol-1-ylmethyl)pentan-3-ol (3 entities in total)
• 機能のキーワード: cyp51, s-tebuconazole, oxidoreductase-oxidoreductase inhibitor complex, oxidoreductase-oxidoreducatse inhibitor complex, oxidoreductase/oxidoreducatse inhibitor
• 由来する生物種: Saccharomyces cerevisiae (Baker's yeast)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 62809.52

構造登録者

Tyndall, J.D.A.,Sabherwal, M.,Sagatova, A.A.,Keniya, M.V.,Wilson, R.K.,Woods, M.V.,Monk, B.C. (登録日: 2015-10-16, 公開日: 2016-02-10, 最終更新日: 2023-09-27)

主引用文献

Tyndall, J.D.,Sabherwal, M.,Sagatova, A.A.,Keniya, M.V.,Negroni, J.,Wilson, R.K.,Woods, M.A.,Tietjen, K.,Monk, B.C.
Structural and Functional Elucidation of Yeast Lanosterol 14 alpha-Demethylase in Complex with Agrochemical Antifungals.
PLoS ONE, 11:e0167485-e0167485, 2016

PubMed Abstract: Azole antifungals, known as demethylase inhibitors (DMIs), target sterol 14α-demethylase (CYP51) in the ergosterol biosynthetic pathway of fungal pathogens of both plants and humans. DMIs remain the treatment of choice in crop protection against a wide range of fungal phytopathogens that have the potential to reduce crop yields and threaten food security. We used a yeast membrane protein expression system to overexpress recombinant hexahistidine-tagged S. cerevisiae lanosterol 14α-demethylase and the Y140F or Y140H mutants of this enzyme as surrogates in order characterize interactions with DMIs. The whole-cell antifungal activity (MIC50 values) of both the R- and S-enantiomers of tebuconazole, prothioconazole (PTZ), prothioconazole-desthio, and oxo-prothioconazole (oxo-PTZ) as well as for fluquinconazole, prochloraz and a racemic mixture of difenoconazole were determined. In vitro binding studies with the affinity purified enzyme were used to show tight type II binding to the yeast enzyme for all compounds tested except PTZ and oxo-PTZ. High resolution X-ray crystal structures of ScErg11p6×His in complex with seven DMIs, including four enantiomers, reveal triazole-mediated coordination of all compounds and the specific orientation of compounds within the relatively hydrophobic binding site. Comparison with CYP51 structures from fungal pathogens including Candida albicans, Candida glabrata and Aspergillus fumigatus provides strong evidence for a highly conserved CYP51 structure including the drug binding site. The structures obtained using S. cerevisiae lanosterol 14α-demethylase in complex with these agrochemicals provide the basis for understanding the impact of mutations on azole susceptibility and a platform for the structure-directed design of the next-generation of DMIs.

PubMed: 27907120
DOI: 10.1371/journal.pone.0167485

実験手法

X-RAY DIFFRACTION (2.6 Å)