9RD1

OppA from E. coli in complex with GSisoK

これはPDB形式変換不可エントリーです。

9RD1 の概要

• エントリーDOI: 10.2210/pdb9rd1/pdb
• 関連するPDBエントリー: 3TCF
• 分子名称: Periplasmic oligopeptide-binding protein OppA, Gly-Ser-epsilon-Lys, (R,R)-2,3-BUTANEDIOL, ... (6 entities in total)
• 機能のキーワード: substrate recognition specificity, binding pocket plasticity, transporter-ligand interactions, structural basis for selective uptake, scaffold accommodation, transport protein
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 238002.86

構造登録者

Iype, T.,Fottner, M.,Boehm, P.,Piedrafita, C.,Moeller, Y.,Groll, M.,Lang, K. (登録日: 2025-05-30, 公開日: 2025-08-13, 最終更新日: 2025-12-10)

主引用文献

Iype, T.,Fottner, M.,Bohm, P.,Piedrafita, C.,Moller, Y.,Groll, M.,Lang, K.
Hijacking a bacterial ABC transporter for genetic code expansion.
Nature, 647:1045-1053, 2025

PubMed Abstract: The site-specific encoding of non-canonical amino acids (ncAAs) provides a powerful tool for expanding the functional repertoire of proteins. Its widespread use for basic research and biotechnological applications is, however, hampered by the low efficiencies of current ncAA incorporation strategies. Here we reveal poor cellular ncAA uptake as a main obstacle to efficient genetic code expansion and overcome this bottleneck by hijacking a bacterial ATP-binding cassette (ABC) transporter to actively import easily synthesizable isopeptide-linked tripeptides that are processed into ncAAs within the cell. Using this approach, we enable efficient encoding of a variety of previously inaccessible ncAAs, decorating proteins with bioorthogonal and crosslinker moieties, post-translational modifications and functionalities for chemoenzymatic conjugation. We then devise a high-throughput directed evolution platform to engineer tailored transporter systems for the import of ncAAs that were historically refractory to efficient uptake. Customized Escherichia coli strains expressing these evolved transporters facilitate single and multi-site ncAA incorporation with wild-type efficiencies. Additionally, we adapt the tripeptide scaffolds for the co-transport of two different ncAAs, enabling their efficient dual incorporation. Collectively, our study demonstrates that engineering of uptake systems is a powerful strategy for programmable import of chemically diverse building blocks.

PubMed: 41094137
DOI: 10.1038/s41586-025-09576-w

実験手法

X-RAY DIFFRACTION (2.5 Å)