9HJV

Crystal structure of flavin-dependent monooxygenase Tet(X4) in complex with prochlorperazine

9HJV の概要

• エントリーDOI: 10.2210/pdb9hjv/pdb
• 分子名称: Flavin-dependent monooxygenase, DIHYDROFLAVINE-ADENINE DINUCLEOTIDE, 2-chloro-10-[3-(4-methylpiperazin-1-yl)propyl]-10H-phenothiazine, ... (5 entities in total)
• 機能のキーワード: flavin-dependent monooxygenase tet(x4), oxidoreductase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 43118.69

構造登録者

Beech, M.J.,Smith, H.G.,Allen, M.D.,Farley, A.J.M.,Schofield, C.J. (登録日: 2024-12-02, 公開日: 2025-05-21, 最終更新日: 2025-06-18)

主引用文献

Beech, M.J.,Toma, E.C.,Smith, H.G.,Trush, M.M.,Ang, J.H.J.,Wong, M.Y.,Wong, C.H.J.,Ali, H.S.,Butt, Z.,Goel, V.,Duarte, F.,Farley, A.J.M.,Walsh, T.R.,Schofield, C.J.
Binding assays enable discovery of Tet(X) inhibitors that combat tetracycline destructase resistance.
Chem Sci, 16:9691-9704, 2025

PubMed Abstract: The Tet(X) flavin-dependent monooxygenases enable tetracycline antibiotic resistance by catalysing inactivating hydroxylation, so preventing inhibition of bacterial ribosomes. Tet(X) resistance is growing rapidly, threatening the efficacy of important last-resort tetracyclines such as tigecycline. Tet(X) inhibitors have potential to protect tetracyclines in combination therapies, but their discovery has been hampered by lack of high-throughput assays. We report the development of an efficient fluorescence polarisation Tet(X) binding assay employing a tetramethylrhodamine-glycyl-minocycline conjugate that enables inhibitor discovery. The assay was applied to tetracycline substrates and reported inhibitors, providing insight into their binding modes. Screening of a bioactive molecule library identified novel Tet(X) inhibitors, including psychoactive phenothiazine derivatives and the 5-HT agonist tegaserod, the activities of which were validated by turnover assays. Crystallographic studies of Tet(X4)-inhibitor complexes reveal two new inhibitor binding modes, importantly providing evidence for active site binding of Tet(X) inhibitors that do not share structural similarity with tetracycline substrates. In some cases, potentiation of tigecycline activity was observed in bacteria expressing Tet(X4). The combined results provide non-tetracycline scaffolds for development of potent Tet(X) inhibitors and highlight the need to evaluate the impact of non-antibiotics on antimicrobial resistance.

PubMed: 40342919
DOI: 10.1039/d5sc00964b

実験手法

X-RAY DIFFRACTION (2.2 Å)