6U0P

Crystal structure of PieE, the flavin-dependent monooxygenase involved in the biosynthesis of piericidin A1

6U0P の概要

• エントリーDOI: 10.2210/pdb6u0p/pdb
• 分子名称: 2,4-dichlorophenol 6-monooxygenase, FLAVIN-ADENINE DINUCLEOTIDE, TRIETHYLENE GLYCOL, ... (7 entities in total)
• 機能のキーワード: flavin-dependent monooxygenase, flavoprotein
• 由来する生物種: Streptomyces sp. SCSIO 03032
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 398391.78

構造登録者

Shi, R.,Manenda, M.,Picard, M.-E. (登録日: 2019-08-14, 公開日: 2020-03-11, 最終更新日: 2023-10-11)

主引用文献

Manenda, M.S.,Picard, M.E.,Zhang, L.,Cyr, N.,Zhu, X.,Barma, J.,Pascal, J.M.,Couture, M.,Zhang, C.,Shi, R.
Structural analyses of the Group A flavin-dependent monooxygenase PieE reveal a sliding FAD cofactor conformation bridging OUT and IN conformations.
J.Biol.Chem., 295:4709-4722, 2020

PubMed Abstract: Group A flavin-dependent monooxygenases catalyze the cleavage of the oxygen-oxygen bond of dioxygen, followed by the incorporation of one oxygen atom into the substrate molecule with the aid of NADPH and FAD. These flavoenzymes play an important role in many biological processes, and their most distinct structural feature is the choreographed motions of flavin, which typically adopts two distinct conformations (OUT and IN) to fulfill its function. Notably, these enzymes seem to have evolved a delicate control system to avoid the futile cycle of NADPH oxidation and FAD reduction in the absence of substrate, but the molecular basis of this system remains elusive. Using protein crystallography, size-exclusion chromatography coupled to multi-angle light scattering (SEC-MALS), and small-angle X-ray scattering (SEC-SAXS) and activity assay, we report here a structural and biochemical characterization of PieE, a member of the Group A flavin-dependent monooxygenases involved in the biosynthesis of the antibiotic piericidin A1. This analysis revealed that PieE forms a unique hexamer. Moreover, we found, to the best of our knowledge for the first time, that in addition to the classical OUT and IN conformations, FAD possesses a "sliding" conformation that exists in between the OUT and IN conformations. This observation sheds light on the underlying mechanism of how the signal of substrate binding is transmitted to the FAD-binding site to efficiently initiate NADPH binding and FAD reduction. Our findings bridge a gap currently missing in the orchestrated order of chemical events catalyzed by this important class of enzymes.

PubMed: 32111738
DOI: 10.1074/jbc.RA119.011212

実験手法

X-RAY DIFFRACTION (2.02 Å)