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

Plug&Pull-In：调整TolC多肽以克服抗生素耐药性 摘要 抗药性感染每年导致的死亡人数已经超过了艾滋病毒/艾滋病或疟疾。临床抗菌药物 耐药性与特定外排泵的过度表达有关。这些外排泵穿梭而出 大多数种类的抗生素使抗生素达不到目标，降低了它们的有效性。这个 外排泵最容易接近的部件是它的外膜桶。外膜桶 可访问性使其成为制药开发的良好目标，因为它可以在没有 穿过通常不透水的外膜。外排泵既是一个可能的目标 用于阻止抗生素外流和一种可能的药物输送途径。堵塞外排泵会使 我们已经拥有的抗生素就像新的一样，通过阻止抗生素从细胞中移除而发挥作用 使他们能够到达目标。劫持桶进行细胞进口可能被用来运送 否则不能穿过细菌外膜。我们的长期目标是开发多肽来对抗 抗生素耐药感染。这项拟议的研究的目的是确定1)什么蛋白质 相互作用是创建高亲和力插头所必需的，以及2)蛋白质-蛋白质相互作用可以促进什么 多肽通过泵传递。我们最近解决了外源蛋白结合的第一个结构 耐药的大肠杆菌的外膜桶。这是一种不同寻常的桶中螺旋结构。我们 已经证明，这种蛋白的结合具有一定的塞子作用。它会阻止外流并使 抗生素的效力更强。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，我们建议利用这些过去的成功来1) 确定抗生素外排泵样蛋白的共同序列和结构桶-肽相互作用2) 筛选与大肠杆菌抗生素外排泵结合良好的多肽，并鉴定决定 这种结合，以及3)确定什么将促进蛋白质在外排泵上的转移， 否则，将其绑定到排气泵。这项工作的全球概念，创造新的插头和移位器 据我们所知，膜β-Barrel以前从未被做过。这些措施的预期结果是什么 实验是关于蛋白质-蛋白质相互作用如何促进堵塞和通过的知识 抗生素外排泵，以及产生堵塞或拉动TolC的多肽。这项工作已经准备就绪 做出重大贡献，因为它将展示重要的蛋白质-蛋白质相互作用 产生膜蛋白塞子和转运体所必需的，从而能够使现有的 对抗耐药超级细菌的抗生素。

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.