6ZMA

Structure of the tRNA-Monooxygenase enzyme MiaE frozen under 140 bar of krypton using the soak and freeze methodology

6ZMA の概要

• エントリーDOI: 10.2210/pdb6zma/pdb
• 分子名称: tRNA hydroxylase, FE (III) ION, CHLORIDE ION, ... (10 entities in total)
• 機能のキーワード: trna-monooxygenase metallo-enzyme trna-modifying enzyme hydroxylase, oxidoreductase
• 由来する生物種: Pseudomonas putida
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 48446.67

構造登録者

Carpentier, P.,Atta, M. (登録日: 2020-07-02, 公開日: 2020-09-02, 最終更新日: 2024-01-31)

主引用文献

Carpentier, P.,Lepretre, C.,Basset, C.,Douki, T.,Torelli, S.,Duarte, V.,Hamdane, D.,Fontecave, M.,Atta, M.
Structural, biochemical and functional analyses of tRNA-monooxygenase enzyme MiaE from Pseudomonas putida provide insights into tRNA/MiaE interaction.
Nucleic Acids Res., 48:9918-9930, 2020

PubMed Abstract: MiaE (2-methylthio-N6-isopentenyl-adenosine37-tRNA monooxygenase) is a unique non-heme diiron enzyme that catalyzes the O2-dependent post-transcriptional allylic hydroxylation of a hypermodified nucleotide 2-methylthio-N6-isopentenyl-adenosine (ms2i6A37) at position 37 of selected tRNA molecules to produce 2-methylthio-N6-4-hydroxyisopentenyl-adenosine (ms2io6A37). Here, we report the in vivo activity, biochemical, spectroscopic characterization and X-ray crystal structure of MiaE from Pseudomonas putida. The investigation demonstrates that the putative pp-2188 gene encodes a MiaE enzyme. The structure shows that Pp-MiaE consists of a catalytic diiron(III) domain with a four alpha-helix bundle fold. A docking model of Pp-MiaE in complex with tRNA, combined with site directed mutagenesis and in vivo activity shed light on the importance of an additional linker region for substrate tRNA recognition. Finally, krypton-pressurized Pp-MiaE experiments, revealed the presence of defined O2 site along a conserved hydrophobic tunnel leading to the diiron active center.

PubMed: 32785618
DOI: 10.1093/nar/gkaa667

実験手法

X-RAY DIFFRACTION (2.15 Å)