6LOI

Crystal structure of Enterococcus faecalis Undecaprenyl pyrophosphate synthase(EfaUPPS)

6LOI の概要

• エントリーDOI: 10.2210/pdb6loi/pdb
• 分子名称: Isoprenyl transferase (2 entities in total)
• 機能のキーワード: isoprenyl transferase, transferase
• 由来する生物種: Enterococcus faecalis
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 200067.94

構造登録者

Lin, W.,Wang, C.Y.,Li, W.J.,Wang, F.L. (登録日: 2020-01-05, 公開日: 2020-06-17, 最終更新日: 2023-11-29)

主引用文献

Li, W.,Li, L.,Zhang, C.,Cai, Y.,Gao, Q.,Wang, F.,Cao, Y.,Lin, J.,Li, J.,Shang, Z.,Lin, W.
Investigations into the Antibacterial Mechanism of Action of Viridicatumtoxins.
Acs Infect Dis., 6:1759-1769, 2020

PubMed Abstract: Viridicatumtoxins are a rare class of tetracycline-like antibiotics that strongly inhibit drug-resistant Gram-positive bacteria. Although reported to exhibit inhibition activity to undecaprenyl pyrophosphate synthase (UPPS), an essential enzyme in bacterial cell wall synthesis, the biological targets and mechanism of action of viridicatumtoxins, especially the drug-target interactions, remain largely unknown. In this study, the structure of UPPS (UPPS) was first determined, uncovering that UPPS can form not only a typical functional dimer but also an unexpected atypical dimer. We then observed that viridicatumtoxins A (VirA) and B (VirB) are able to bind to UPPSs of , , and in a direct and high-affinity manner as evidenced by enzyme inhibition assay, surface plasmon resonance (SPR) binding analysis, and growth inhibition assay, demonstrating that viridicatumtoxins exert antibacterial effects through UPPS binding. The key amino acid residues involved in the interactions with VirA and VirB in UPPS binding pocket were revealed by molecular docking studies, and further validated by site-directed mutagenesis. A single mutation of UPPS at D29A, N31A, and R42A can obviously increase their affinities to VirA, while a single mutation at W228A conferred significant resistance to VirA. Moreover, translation inhibition assay showed that VirA and VirB can weakly inhibit 70S ribosome. The weak inhibition of ribosome was proposed to be attributed to steric hindrance between viridicatumtoxin ring F and 70S ribosome helix 34 by molecular docking study. Our structural, biochemical, and computational investigations on the interactions of viridicatumtoxins with UPPS and 70S ribosome not only disclosed the potential biological targets of viridicatumtoxins, but also provided a theoretical basis for structural optimization to make new viridicatumtoxin derivatives with improved antimicrobial activities.

PubMed: 32437130
DOI: 10.1021/acsinfecdis.0c00031

実験手法

X-RAY DIFFRACTION (2.503 Å)