8ERJ

Crystal structure of Fub7 in complex with E-2-aminocrotonate

8ERJ の概要

• エントリーDOI: 10.2210/pdb8erj/pdb
• 分子名称: Sulfhydrylase FUB7, (2E)-2-{[(1E)-{3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl}methylidene]amino}but-2-enoic acid, (2S)-2-[({3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl}methyl)amino]but-3-enoic acid, ... (4 entities in total)
• 機能のキーワード: c-c bond formation, biosynthetic protein
• 由来する生物種: Fusarium fujikuroi
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 396549.84

構造登録者

Hai, Y. (登録日: 2022-10-12, 公開日: 2023-10-25, 最終更新日: 2024-11-06)

主引用文献

Liu, S.,Yeh, C.,Reavill, C.,Jones, B.,Zou, Y.,Hai, Y.
Molecular and Structural Basis for C gamma-C Bond Formation by PLP-Dependent Enzyme Fub7.
Angew.Chem.Int.Ed.Engl., 63:e202317161-e202317161, 2024

PubMed Abstract: Pyridoxal 5'-phosphate (PLP)-dependent enzymes that catalyze γ-replacement reactions are prevalent, yet their utilization of carbon nucleophile substrates is rare. The recent discovery of two PLP-dependent enzymes, CndF and Fub7, has unveiled unique C-C bond forming capabilities, enabling the biocatalytic synthesis of alkyl- substituted pipecolic acids from O-acetyl-L-homoserine and β-keto acid or aldehyde derived enolates. This breakthrough presents fresh avenues for the biosynthesis of pipecolic acid derivatives. However, the catalytic mechanisms of these enzymes remain elusive, and a dearth of structural information hampers their extensive application. Here, we have broadened the catalytic scope of Fub7 by employing ketone-derived enolates as carbon nucleophiles, revealing Fub7's capacity for substrate-dependent regioselective α-alkylation of unsymmetrical ketones. Through an integrated approach combining X-ray crystallography, spectroscopy, mutagenesis, and computational docking studies, we offer a detailed mechanistic insight into Fub7 catalysis. Our findings elucidate the structural basis for its substrate specificity, stereoselectivity, and regioselectivity. Our work sets the stage ready for subsequent protein engineering effort aimed at expanding the synthetic utility of Fub7, potentially unlocking novel methods to access a broader array of noncanonical amino acids.

PubMed: 38308582
DOI: 10.1002/anie.202317161

実験手法

X-RAY DIFFRACTION (2.16 Å)