8YYR

Structure of the HitB T293G mutant

これはPDB形式変換不可エントリーです。

8YYR の概要

• エントリーDOI: 10.2210/pdb8yyr/pdb
• 分子名称: Putative ATP-dependent b-aminoacyl-ACP synthetase, [(2~{R},3~{S},4~{R},5~{R})-5-(6-aminopurin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methyl ~{N}-[(3~{S})-3-azanyl-3-(2-bromophenyl)propanoyl]sulfamate (3 entities in total)
• 機能のキーワード: hitachimycin, polyketide biosynthesis, atp binding, adenylation, ligase
• 由来する生物種: Embleya scabrispora
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 60203.04

構造登録者

Wang, D.,Miyanaga, A.,Chisuga, T.,Kudo, F.,Eguchi, T. (登録日: 2024-04-04, 公開日: 2024-06-05, 最終更新日: 2024-08-14)

主引用文献

Wang, D.,Miyanaga, A.,Chisuga, T.,Kudo, F.,Eguchi, T.
Engineering the Substrate Specificity of (S)-beta-Phenylalanine Adenylation Enzyme HitB.
Chembiochem, 25:e202400383-e202400383, 2024

PubMed Abstract: Adenylation enzymes catalyze the selective incorporation of aminoacyl building blocks in the biosynthesis of nonribosomal peptides and related natural products. Although β-amino acid units are one of the important aminoacyl building blocks in natural product biosynthesis, very little is known about the engineering of β-amino acid adenylation enzymes. In this study, we engineered the substrate specificity of the (S)-β-phenylalanine adenylation enzyme, HitB, involved in the biosynthesis of macrolactam polyketide hitachimycin. Based on the previously determined structure of HitB wild-type, we mutated Phe328 and Ser293, which are located near the meta and ortho position of the (S)-β-phenylalanine moiety, respectively. As a result, the HitB F328V and F328L mutants efficiently activated meta-substituted (S)-β-phenylalanine analogs, and the HitB T293G and T293S mutants efficiently activated ortho-substituted (S)-β-phenylalanine analogs. Structural analysis of the HitB F328L and T293G mutants with the corresponding nonhydrolyzable intermediate analogs revealed an enlarged substrate binding pocket for (S)-β-phenylalanine analogs, providing detailed insights into the structural basis for creating enzyme substrate promiscuity. Our findings may be useful for production of various β-amino acid-containing natural product analogs.

PubMed: 38805007
DOI: 10.1002/cbic.202400383

実験手法

X-RAY DIFFRACTION (1.8 Å)