6MAX

Crystal structure of Ribonuclease P protein from Thermotoga maritima in complex with purpurin

6MAX の概要

• エントリーDOI: 10.2210/pdb6max/pdb
• 関連するPDBエントリー: 1NZ0
• 分子名称: Ribonuclease P protein component, SULFATE ION, Purpurin, ... (4 entities in total)
• 機能のキーワード: p protein, rna binding protein, hydrolase
• 由来する生物種: Thermotoga maritima
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14272.77

構造登録者

Torres-Larios, A.,Madrigal-Carrillo, E.A. (登録日: 2018-08-28, 公開日: 2019-04-17, 最終更新日: 2023-10-11)

主引用文献

Madrigal-Carrillo, E.A.,Diaz-Tufinio, C.A.,Santamaria-Suarez, H.A.,Arciniega, M.,Torres-Larios, A.
A screening platform to monitor RNA processing and protein-RNA interactions in ribonuclease P uncovers a small molecule inhibitor.
Nucleic Acids Res., 47:6425-6438, 2019

PubMed Abstract: Ribonucleoprotein (RNP) complexes and RNA-processing enzymes are attractive targets for antibiotic development owing to their central roles in microbial physiology. For many of these complexes, comprehensive strategies to identify inhibitors are either lacking or suffer from substantial technical limitations. Here, we describe an activity-binding-structure platform for bacterial ribonuclease P (RNase P), an essential RNP ribozyme involved in 5' tRNA processing. A novel, real-time fluorescence-based assay was used to monitor RNase P activity and rapidly identify inhibitors using a mini-helix and a pre-tRNA-like bipartite substrate. Using the mini-helix substrate, we screened a library comprising 2560 compounds. Initial hits were then validated using pre-tRNA and the pre-tRNA-like substrate, which ultimately verified four compounds as inhibitors. Biolayer interferometry-based binding assays and molecular dynamics simulations were then used to characterize the interactions between each validated inhibitor and the P protein, P RNA and pre-tRNA. X-ray crystallographic studies subsequently elucidated the structure of the P protein bound to the most promising hit, purpurin, and revealed how this inhibitor adversely affects tRNA 5' leader binding. This integrated platform affords improved structure-function studies of RNA processing enzymes and facilitates the discovery of novel regulators or inhibitors.

PubMed: 30997498
DOI: 10.1093/nar/gkz285

実験手法

X-RAY DIFFRACTION (1.42 Å)