6YA5

2009 H1N1 PA Endonuclease in complex with LU2

6YA5 の概要

• エントリーDOI: 10.2210/pdb6ya5/pdb
• 分子名称: Polymerase acidic protein,Polymerase acidic protein, MANGANESE (II) ION, MAGNESIUM ION, ... (9 entities in total)
• 機能のキーワード: influenza, polymerase, endonuclease, hydrolase, viral protein
• 由来する生物種: Influenza A virus (A/California/07/2009(H1N1)); 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 22013.13

構造登録者

Radilova, K.,Brynda, J. (登録日: 2020-03-11, 公開日: 2020-09-09, 最終更新日: 2024-01-24)

主引用文献

Zima, V.,Radilova, K.,Kozisek, M.,Albinana, C.B.,Karlukova, E.,Brynda, J.,Fanfrlik, J.,Flieger, M.,Hodek, J.,Weber, J.,Majer, P.,Konvalinka, J.,Machara, A.
Unraveling the anti-influenza effect of flavonoids: Experimental validation of luteolin and its congeners as potent influenza endonuclease inhibitors.
Eur.J.Med.Chem., 208:112754-112754, 2020

PubMed Abstract: The biological effects of flavonoids on mammal cells are diverse, ranging from scavenging free radicals and anti-cancer activity to anti-influenza activity. Despite appreciable effort to understand the anti-influenza activity of flavonoids, there is no clear consensus about their precise mode-of-action at a cellular level. Here, we report the development and validation of a screening assay based on AlphaScreen technology and illustrate its application for determination of the inhibitory potency of a large set of polyols against PA N-terminal domain (PA-Nter) of influenza RNA-dependent RNA polymerase featuring endonuclease activity. The most potent inhibitors we identified were luteolin with an IC of 72 ± 2 nM and its 8-C-glucoside orientin with an IC of 43 ± 2 nM. Submicromolar inhibitors were also evaluated by an in vitro endonuclease activity assay using single-stranded DNA, and the results were in full agreement with data from the competitive AlphaScreen assay. Using X-ray crystallography, we analyzed structures of the PA-Nter in complex with luteolin at 2.0 Å resolution and quambalarine B at 2.5 Å resolution, which clearly revealed the binding pose of these polyols coordinated to two manganese ions in the endonuclease active site. Using two distinct assays along with the structural work, we have presumably identified and characterized the molecular mode-of-action of flavonoids in influenza-infected cells.

PubMed: 32883638
DOI: 10.1016/j.ejmech.2020.112754

実験手法

X-RAY DIFFRACTION (2 Å)