6VHY

NpsA-ThdA, an artificially fused Adenylation-PCP di-domain NRPS from Klebsiella oxytoca

6VHY の概要

• エントリーDOI: 10.2210/pdb6vhy/pdb
• 分子名称: NpsA Adenylation Domain, Non-ribosomal peptide synthetase fusion protein, 5'-deoxy-5'-({[(2R)-2-{[2-({N-[(2R)-2-hydroxy-3,3-dimethyl-4-(phosphonooxy)butanoyl]-beta-alanyl}amino)ethyl]sulfanyl}-2-(3-hydroxyphenyl)ethyl]sulfonyl}amino)adenosine (3 entities in total)
• 機能のキーワード: adenylation, tilivalline, tilimycin, nrps, nonribosomal peptide synthetase, biosynthetic protein
• 由来する生物種: Klebsiella oxytoca; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 259477.30

構造登録者

Kreitler, D.F.,Gulick, A.M. (登録日: 2020-01-10, 公開日: 2020-06-24, 最終更新日: 2023-10-11)

主引用文献

Alexander, E.M.,Kreitler, D.F.,Guidolin, V.,Hurben, A.K.,Drake, E.,Villalta, P.W.,Balbo, S.,Gulick, A.M.,Aldrich, C.C.
Biosynthesis, Mechanism of Action, and Inhibition of the Enterotoxin Tilimycin Produced by the Opportunistic PathogenKlebsiella oxytoca.
Acs Infect Dis., 6:1976-1997, 2020

PubMed Abstract: Tilimycin is an enterotoxin produced by the opportunistic pathogen that causes antibiotic-associated hemorrhagic colitis (AAHC). This pyrrolobenzodiazepine (PBD) natural product is synthesized by a bimodular nonribosomal peptide synthetase (NRPS) pathway composed of three proteins: NpsA, ThdA, and NpsB. We describe the functional and structural characterization of the fully reconstituted NRPS system and report the steady-state kinetic analysis of all natural substrates and cofactors as well as the structural characterization of both NpsA and ThdA. The mechanism of action of tilimycin was confirmed using DNA adductomics techniques through the detection of putative N-2 guanine alkylation after tilimycin exposure to eukaryotic cells, providing the first structural characterization of a PBD-DNA adduct formed in cells. Finally, we report the rational design of small-molecule inhibitors that block tilimycin biosynthesis in whole cell (IC = 29 ± 4 μM) through the inhibition of NpsA ( = 29 ± 4 nM).

PubMed: 32485104
DOI: 10.1021/acsinfecdis.0c00326

実験手法

X-RAY DIFFRACTION (3 Å)