Plugging & Pulling-in: tuning peptides for ToIC to overcome anitbiotic resistance

堵漏

• 批准号: 10737465
• 负责人: Joanna SG Slusky
• 金额: $ 31.12万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-05 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737465
• 关键词: Affinity; Amino Acids; Aminoglycosides; Antibiotic Resistance; Antibiotics; Architecture; Bacteria; Bacterial Translocation; Binding; Binding Proteins; Biological Assay; Calorimetry; Cells; Cessation of life; Chemosensitization; Chloramphenicol; Clinical; Data Set; Databases; Development; Drug Delivery Systems; E coli tolC protein; Effectiveness; Escherichia coli; Evolution; Fluorescence; Fluoroquinolones; Frequencies; Geometry; Goals; Gram-Negative Bacteria; HIV/AIDS; Infection; Knowledge; Length; Libraries; Macrolides; Malaria; Measures; Membrane; Membrane Proteins; Outcome; Pattern; Peptide Library; Peptides; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Positioning Attribute; Predisposition; Process; Protein Import; Protein translocation; Proteins; Pump; Research; Resistance; Structure; Superbug; Surface; System; Tetracyclines; Time; Titrations; Triclosan; Visualization; Work; Yeasts; alpha helix; antibiotic efflux; antibiotic resistant infections; beta barrel; beta-Lactams; colicin; combat; efflux pump; experimental study; fighting; global health; improved; interest; novel therapeutics; overexpression; protein protein interaction; prototype; quaternary ammonium compound; single molecule; success; tool

Plugging & Pulling-in: tuning peptides for TolC to overcome antibiotic resistance Summary Antibiotic resistant infections already kill more people per year than HIV/AIDS or malaria. Clinical antibiotic resistance is correlated with the overexpression of particular efflux pumps. These efflux pumps shuttle out most classes of antibiotics so that the antibiotics can’t reach their targets, reducing their effectiveness. The most accessible part of the efflux pump is its outer membrane barrel. The outer membrane barrel accessibility makes it a good target for pharmaceutical development because it can be accessed without getting through the often impermeable outer membrane. The efflux pump presents as both a possible target for stopping antibiotic efflux and a possible avenue of drug delivery. Plugging efflux pumps would make the antibiotics we already have work like new by stopping antibiotics from being removed from the cell and allowing them to reach their targets. Hijacking the barrel for cellular import could be used to deliver drugs that otherwise could not cross the bacterial outer membrane. Our long term goal is to develop peptides to combat antibiotic resistant infection. The objective of the proposed research is to determine 1) what protein-protein interactions are necessary to create high affinity plugs and 2) what protein-protein interactions can facilitate peptide transit through the pump. We have recently solved the first structure of an exogenous protein binding the outer membrane barrel of the E. coli antibiotic resistance. It is an unusual helix-in-barrel structure. We have shown that the binding of this protein has some ability to act as a plug. It stops efflux and make antibiotics more potent. We have also created databases of outer membrane barrels to better understand the sequence structure relationships in these proteins. Here we propose to leverage these past successes to 1) Identify common sequence and structural barrel-peptide interactions for antibiotic efflux pump-like proteins 2) Screen for peptides that bind well to the E. coli antibiotic efflux pump and identify the features that determine that binding, and 3) Identify what would facilitate the translocation of proteins across the efflux pump that otherwise bind to the efflux pump. The global concept of this work, creating new plugs and translocators for membrane β-barrels, has, to our knowledge, never been done before. The expected outcomes of these experiments are a knowledge of how protein-protein interactions facilitate plugging and pulling through of antibiotic efflux pumps, as well as the creation of peptides that plug or pull through TolC. This work is poised to make a significant contribution because it will demonstrate the important protein-protein interactions necessary to create membrane protein plugs and translocators and can thereby enable a revival of existing antibiotics against resistant superbugs.

插入和拉入：TOLC的调整肽以克服抗生素耐药性 概括 抗生素耐药性感染每年已经杀死的人数比艾滋病毒/艾滋病或疟疾更多。临床抗生素 电阻与特定外排泵的过表达相关。这些外排泵驶出 大多数类别的抗生素，因此抗生素无法达到其靶标，从而降低了其有效性。这 外排泵最容易接近的部分是其外膜枪管。外膜桶 可访问性使其成为药物开发的好目标，因为可以在没有的情况下访问 通过通常不可渗透的外膜。排出泵既是可能的目标 用于停止抗生素外排和可能的药物输送途径。插入外泵将使 抗生素我们已经通过阻止抗生素从细胞中去除和 允许他们达到目标。劫持枪管的蜂窝进口可以用来输送药物 否则无法越过细菌外膜。我们的长期目标是开发肽来对抗 抗生素抗性感染。拟议研究的目的是确定1）哪种蛋白质蛋白 相互作用对于创建高亲和力插头是必要的，2）哪些蛋白质 - 蛋白质相互作用可以促进 肽通过泵的转运。我们最近解决了外源蛋白结合的第一个结构 大肠杆菌抗生素耐药性的外膜桶。这是一种不寻常的螺旋内桶结构。我们 已经表明，该蛋白质的结合具有某种充当插头的能力。它停止排出并制造 抗生素的潜力更大。我们还创建了外膜桶的数据库，以更好地了解 这些蛋白质中的序列结构关系。在这里，我们建议将过去的成功利用1） 确定用于抗生素外排泵样蛋白的常见序列和结构桶肽相互作用2） 屏幕上的辣椒与大肠杆菌抗生素外排泵结合良好并确定确定的特征 结合，3）确定什么将有助于蛋白质在外排泵上的易位 否则与外排泵结合。这项工作的全球概念，为 据我们所知，膜β桶从未做过。这些预期的结果 实验是了解蛋白质 - 蛋白质相互作用如何促进和拉动的知识 抗生素外排泵，以及插入或拉动TOLC的Petides的创建。这项工作被中毒 做出重大贡献，因为它将证明重要的蛋白质 - 蛋白质相互作用 创建膜蛋白插头和转运器所需的必要条件，从而使现有的复兴 抵抗超级细菌的抗生素。