5L86

engineered ascorbate peroxidise

5L86 の概要

• エントリーDOI: 10.2210/pdb5l86/pdb
• 分子名称: Ascorbate peroxidase, PROTOPORPHYRIN IX CONTAINING FE, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: protein engineering, peroxidase, heme, apx2, methyl histidine, oxidoreductase
• 由来する生物種: Glycine max (Soybean)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 55437.82

構造登録者

Hayashi, T.,Mittl, P.,Hilvert, D. (登録日: 2016-06-07, 公開日: 2017-03-01, 最終更新日: 2024-01-10)

主引用文献

Green, A.P.,Hayashi, T.,Mittl, P.R.,Hilvert, D.
A Chemically Programmed Proximal Ligand Enhances the Catalytic Properties of a Heme Enzyme.
J. Am. Chem. Soc., 138:11344-11352, 2016

PubMed Abstract: Enzymes rely on complex interactions between precisely positioned active site residues as a mechanism to compensate for the limited functionality contained within the genetic code. Heme enzymes provide a striking example of this complexity, whereby the electronic properties of reactive ferryl intermediates are finely tuned through hydrogen bonding interactions between proximal ligands and neighboring amino acids. Here, we show that introduction of a chemically programmed proximal Nδ-methyl histidine (NMH) ligand into an engineered ascorbate peroxidase (APX2) overcomes the reliance on the conserved Asp-His hydrogen bonding interaction, leading to a catalytically modified enzyme (APX2 NMH), which is able to achieve a significantly higher number of turnovers compared with APX2 without compromising catalytic efficiency. Structural, spectroscopic and kinetic characterization of APX2 NMH and several active site variants provides valuable insights into the role of the Asp-His-Fe triad of heme peroxidases. More significantly, simplification of catalytic mechanisms through the incorporation of chemically optimized ligands may facilitate efforts to create and evolve new active site heme environments within proteins.

PubMed: 27500802
DOI: 10.1021/jacs.6b07029

実験手法

X-RAY DIFFRACTION (1.9 Å)