7YAV

Crystal structure of Diels-Alderase MaDA1

7YAV の概要

• エントリーDOI: 10.2210/pdb7yav/pdb
• 分子名称: MaDA1, FLAVIN-ADENINE DINUCLEOTIDE, 2-acetamido-2-deoxy-beta-D-glucopyranose, ... (4 entities in total)
• 機能のキーワード: natural product biosynthetic enzyme, plant protein
• 由来する生物種: Morus alba
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 124544.95

構造登録者

Lei, X.G.,Chen, R.C.,Du, X.X.,Yang, J.,Fan, J.P.,Guo, N.X.,Ding, Q. (登録日: 2022-06-28, 公開日: 2024-01-24, 最終更新日: 2026-03-04)

主引用文献

Ding, Q.,Guo, N.,Gao, L.,McKee, M.,Wu, D.,Yang, J.,Fan, J.,Weng, J.K.,Lei, X.
The evolutionary origin of naturally occurring intermolecular Diels-Alderases from Morus alba.
Nat Commun, 15:2492-2492, 2024

PubMed Abstract: Biosynthetic enzymes evolutionarily gain novel functions, thereby expanding the structural diversity of natural products to the benefit of host organisms. Diels-Alderases (DAs), functionally unique enzymes catalysing [4 + 2] cycloaddition reactions, have received considerable research interest. However, their evolutionary mechanisms remain obscure. Here, we investigate the evolutionary origins of the intermolecular DAs in the biosynthesis of Moraceae plant-derived Diels-Alder-type secondary metabolites. Our findings suggest that these DAs have evolved from an ancestor functioning as a flavin adenine dinucleotide (FAD)-dependent oxidocyclase (OC), which catalyses the oxidative cyclisation reactions of isoprenoid-substituted phenolic compounds. Through crystal structure determination, computational calculations, and site-directed mutagenesis experiments, we identified several critical substitutions, including S348L, A357L, D389E and H418R that alter the substrate-binding mode and enable the OCs to gain intermolecular DA activity during evolution. This work provides mechanistic insights into the evolutionary rationale of DAs and paves the way for mining and engineering new DAs from other protein families.

PubMed: 38509059
DOI: 10.1038/s41467-024-46845-0

実験手法

X-RAY DIFFRACTION (2.1 Å)