High-Throughput Identification and Characterization of LptDE-Binding Bicycle Peptides Using Phage Display and Cryo-EM.

IF 6.8 1区医学 Q1 CHEMISTRY, MEDICINAL

Journal of Medicinal Chemistry Pub Date : 2025-10-06 DOI:10.1021/acs.jmedchem.5c00307

Shenaz Allyjaun,Emily Dunbar,Steven W Hardwick,Sarah Newell,Finn Holding,Catherine E Rowland,Megan A St Denis,Simone Pellegrino,Gustavo Arruda Bezerra,Nikolaos Bournakas,Dimitri Y Chirgadze,Lee Cooper,Giulia Paris,Nick Lewis,Peter Brown,Michael J Skynner,Michael J Dawson,Paul Beswick,Julia Hubbard,Bert van den Berg,Hector Newman

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引用次数: 0

Abstract

The lipopolysaccharide (LPS) transport (Lpt) system in Gram-negative bacteria maintains the integrity of the asymmetric bacterial outer membrane (OM). LPS biogenesis systems are essential in most Gram-negative bacteria, with LptDE responsible for the delivery of LPS to the outer leaflet of the OM. As an externally accessible, essential protein, LptDE offers a promising target for inhibitor development without the need for cellular penetration. However, there are no direct inhibitors of E. coli LptDE, and drug discovery is made challenging since it is a membrane target without a conventional active site. Here, the bicycle phage display platform was used in combination with cryogenic-electron microscopy (cryo-EM) and surface plasmon resonance to identify and map bicyclic peptide binders to Shigella flexneri LptDE (SfLptDE). Four distinct epitopes with unique bicycle molecule binding motifs were identified across the SfLptD β-barrel. This method represents a streamlined workflow for the identification and prioritization of hit molecules against LptDE.

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利用噬菌体展示和低温电镜技术高通量鉴定和表征lptde结合的环状肽。

革兰氏阴性菌的脂多糖（LPS）转运（Lpt）系统维持了不对称细菌外膜（OM）的完整性。脂多糖生物生成系统在大多数革兰氏阴性菌中是必不可少的，LptDE负责将脂多糖传递到OM的外层小叶。作为一种外部可及的必需蛋白，LptDE为抑制剂的开发提供了一个有希望的靶点，而不需要细胞渗透。然而，目前还没有大肠杆菌LptDE的直接抑制剂，而且由于它是一种没有常规活性位点的膜靶点，因此药物发现具有挑战性。本研究利用自行车噬菌体展示平台，结合低温电子显微镜（cro - em）和表面等离子体共振技术，鉴定并定位了双环肽结合物与福氏志贺氏杆菌LptDE （SfLptDE）的关系。在SfLptD β-桶中鉴定出四个具有独特的自行车分子结合基序的不同表位。该方法代表了一种简化的工作流程，用于识别和确定针对LptDE的命中分子的优先级。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Medicinal Chemistry 医学-医药化学

CiteScore

4.00

自引率

11.00%

发文量

804

审稿时长

1.9 months

期刊介绍： The Journal of Medicinal Chemistry is a prestigious biweekly peer-reviewed publication that focuses on the multifaceted field of medicinal chemistry. Since its inception in 1959 as the Journal of Medicinal and Pharmaceutical Chemistry, it has evolved to become a cornerstone in the dissemination of research findings related to the design, synthesis, and development of therapeutic agents. The Journal of Medicinal Chemistry is recognized for its significant impact in the scientific community, as evidenced by its 2022 impact factor of 7.3. This metric reflects the journal's influence and the importance of its content in shaping the future of drug discovery and development. The journal serves as a vital resource for chemists, pharmacologists, and other researchers interested in the molecular mechanisms of drug action and the optimization of therapeutic compounds.