6GMI

Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in E. coli's Periplasm.

6GMI の概要

• エントリーDOI: 10.2210/pdb6gmi/pdb
• 分子名称: Streptavidin, {N-(4-{[2-(amino-kappaN)ethyl]sulfamoyl-kappaN}phenyl)-5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamide}(chloro)[(1,2,3,4,5-eta)-1,2,3,4,5-pentamethylcyclopentadienyl]iridium(III), IRIDIUM (III) ION, ... (4 entities in total)
• 機能のキーワード: biotin-binding protein, artificial transfer hydrogenase, beta barrel, streptavidin
• 由来する生物種: Streptomyces avidinii
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 20200.62

構造登録者

Rebelein, J.G. (登録日: 2018-05-26, 公開日: 2018-10-10, 最終更新日: 2024-01-17)

主引用文献

Zhao, J.,Rebelein, J.G.,Mallin, H.,Trindler, C.,Pellizzoni, M.M.,Ward, T.R.
Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coli's Periplasm.
J. Am. Chem. Soc., 140:13171-13175, 2018

PubMed Abstract: Artificial metalloenzymes (ArMs), which combine an abiotic metal cofactor with a protein scaffold, catalyze various synthetically useful transformations. To complement the natural enzymes' repertoire, effective optimization protocols to improve ArM's performance are required. Here we report on our efforts to optimize the activity of an artificial transfer hydrogenase (ATHase) using Escherichia coli whole cells. For this purpose, we rely on a self-immolative quinolinium substrate which, upon reduction, releases fluorescent umbelliferone, thus allowing efficient screening. Introduction of a loop in the immediate proximity of the Ir-cofactor afforded an ArM with up to 5-fold increase in transfer hydrogenation activity compared to the wild-type ATHase using purified mutants.

PubMed: 30272972
DOI: 10.1021/jacs.8b07189

実験手法

X-RAY DIFFRACTION (1.6 Å)