6IYK

The structure of EntE with 2-nitrobenzoyl adenylate analog

6IYK の概要

• エントリーDOI: 10.2210/pdb6iyk/pdb
• 分子名称: 2,3-dihydroxybenzoate-AMP ligase component of enterobactin synthase multienzyme complex, 5'-O-[(2-nitrobenzene-1-carbonyl)sulfamoyl]adenosine (3 entities in total)
• 機能のキーワード: ligase, adenylation, non-ribosomal peptide biosynthesis
• 由来する生物種: Escherichia coli 1303
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 123657.30

構造登録者

Miyanaga, A.,Ishikawa, F. (登録日: 2018-12-17, 公開日: 2019-04-17, 最終更新日: 2023-11-22)

主引用文献

Ishikawa, F.,Miyanaga, A.,Kitayama, H.,Nakamura, S.,Nakanishi, I.,Kudo, F.,Eguchi, T.,Tanabe, G.
An Engineered Aryl Acid Adenylation Domain with an Enlarged Substrate Binding Pocket.
Angew.Chem.Int.Ed.Engl., 58:6906-6910, 2019

PubMed Abstract: Adenylation (A) domains act as the gatekeepers of non-ribosomal peptide synthetases (NRPSs), ensuring the activation and thioesterification of the correct amino acid/aryl acid building blocks. Aryl acid building blocks are most commonly observed in iron-chelating siderophores, but are not limited to them. Very little is known about the reprogramming of aryl acid A-domains. We show that a single asparagine-to-glycine mutation in an aryl acid A-domain leads to an enzyme that tolerates a wide range of non-native aryl acids. The engineered catalyst is capable of activating non-native aryl acids functionalized with nitro, cyano, bromo, and iodo groups, even though no enzymatic activity of wild-type enzyme was observed toward these substrates. Co-crystal structures with non-hydrolysable aryl-AMP analogues revealed the origins of this expansion of substrate promiscuity, highlighting an enlargement of the substrate binding pocket of the enzyme. Our findings may be exploited to produce diversified aryl acid containing natural products and serve as a template for further directed evolution in combinatorial biosynthesis.

PubMed: 30945421
DOI: 10.1002/anie.201900318

実験手法

X-RAY DIFFRACTION (2.45 Å)