7ANV

Mutational and structural analysis of an ancestral D-type dye decolorizing peroxidase

7ANV の概要

• エントリーDOI: 10.2210/pdb7anv/pdb
• 分子名称: ancestral D-type dye decolorizing peroxidase, PROTOPORPHYRIN IX CONTAINING FE, GLYCEROL, ... (6 entities in total)
• 機能のキーワード: dye decolorizing peroxidase, d-type dyp, oxidoreductase
• 由来する生物種: Geotrichum candidum
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 54736.49

構造登録者

Rozeboom, H.J.,Fraaije, M.W. (登録日: 2020-10-12, 公開日: 2020-12-30, 最終更新日: 2024-01-31)

主引用文献

Zitare, U.A.,Habib, M.H.,Rozeboom, H.,Mascotti, M.L.,Todorovic, S.,Fraaije, M.W.
Mutational and structural analysis of an ancestral fungal dye-decolorizing peroxidase.
Febs J., 288:3602-3618, 2021

PubMed Abstract: Dye-decolorizing peroxidases (DyPs) constitute a superfamily of heme-containing peroxidases that are related neither to animal nor to plant peroxidase families. These are divided into four classes (types A, B, C, and D) based on sequence features. The active site of DyPs contains two highly conserved distal ligands, an aspartate and an arginine, the roles of which are still controversial. These ligands have mainly been studied in class A-C bacterial DyPs, largely because no effective recombinant expression systems have been developed for the fungal (D-type) DyPs. In this work, we employ ancestral sequence reconstruction (ASR) to resurrect a D-type DyP ancestor, AncDyPD-b1. Expression of AncDyPD-b1 in Escherichia coli results in large amounts of a heme-containing soluble protein and allows for the first mutagenesis study on the two distal ligands of a fungal DyP. UV-Vis and resonance Raman (RR) spectroscopic analyses, in combination with steady-state kinetics and the crystal structure, reveal fine pH-dependent details about the heme active site structure and show that both the aspartate (D222) and the arginine (R390) are crucial for hydrogen peroxide reduction. Moreover, the data indicate that these two residues play important but mechanistically different roles on the intraprotein long-range electron transfer process. DATABASE: Structural data are available in the PDB database under the accession number 7ANV.

PubMed: 33369202
DOI: 10.1111/febs.15687

実験手法

X-RAY DIFFRACTION (1.65 Å)