5OA4

Fe(II)/(alpha)ketoglutarate-dependent dioxygenase AsqJ_V72I mutant in complex with 4-methoxycyclopeptin (1)

5OA4 の概要

• エントリーDOI: 10.2210/pdb5oa4/pdb
• 関連するPDBエントリー: 5DAP
• 分子名称: Iron/alpha-ketoglutarate-dependent dioxygenase asqJ, NICKEL (II) ION, (3S)-3-(4-methoxybenzyl)-4-methyl-3,4-dihydro-1H-1,4-benzodiazepine-2,5-dione, ... (6 entities in total)
• 機能のキーワード: antibiotics, quinolone biosynthesis, molecular engineering, desaturase, catalytic mechanism, mutagenesis, pi-stacking, oxidoreductase
• 由来する生物種: Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34519.52

構造登録者

Groll, M.,Braeuer, A.,Kaila, V.R.I. (登録日: 2017-06-20, 公開日: 2018-04-04, 最終更新日: 2024-01-17)

主引用文献

Mader, S.L.,Brauer, A.,Groll, M.,Kaila, V.R.I.
Catalytic mechanism and molecular engineering of quinolone biosynthesis in dioxygenase AsqJ.
Nat Commun, 9:1168-1168, 2018

PubMed Abstract: The recently discovered Fe/α-ketoglutarate-dependent dioxygenase AsqJ from Aspergillus nidulans stereoselectively catalyzes a multistep synthesis of quinolone alkaloids, natural products with significant biomedical applications. To probe molecular mechanisms of this elusive catalytic process, we combine here multi-scale quantum and classical molecular simulations with X-ray crystallography, and in vitro biochemical activity studies. We discover that methylation of the substrate is essential for the activity of AsqJ, establishing molecular strain that fine-tunes π-stacking interactions within the active site. To rationally engineer AsqJ for modified substrates, we amplify dispersive interactions within the active site. We demonstrate that the engineered enzyme has a drastically enhanced catalytic activity for non-methylated surrogates, confirming our computational data and resolved high-resolution X-ray structures at 1.55 Å resolution. Our combined findings provide crucial mechanistic understanding of the function of AsqJ and showcase how combination of computational and experimental data enables to rationally engineer enzymes.

PubMed: 29563492
DOI: 10.1038/s41467-018-03442-2

実験手法

X-RAY DIFFRACTION (1.55 Å)