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

Plugging & Pulling-in：调整TolC肽以克服抗生素耐药性 总结 抗生素耐药性感染每年造成的死亡人数已经超过艾滋病毒/艾滋病或疟疾。临床抗菌药物 抗性与特定外排泵的过表达相关。这些外排泵穿梭于 大多数种类的抗生素，使抗生素不能达到他们的目标，降低其有效性。的 外排泵的最容易接近的部分是其外膜筒。外膜筒 可及性使其成为药物开发的良好目标，因为它可以在没有 穿过通常不渗透的外膜。外排泵既是一个可能的目标 用于阻止抗生素外排和可能的药物输送途径。堵塞外排泵会使 我们已有的抗生素像新的一样工作，阻止抗生素从细胞中被清除， 使他们能够达到目标。劫持细胞进口桶可以用来运送药物， 否则不能穿过细菌外膜。我们的长期目标是开发肽来对抗 抗生素耐药性感染拟议研究的目的是确定1）什么蛋白质-蛋白质 相互作用是产生高亲和力栓所必需的，以及2）蛋白质-蛋白质相互作用可以促进什么 肽通过泵转运。我们最近解决了第一个结构的外源蛋白结合 E.大肠杆菌抗生素抗性。这是一个不寻常的螺旋桶结构。我们 已经表明这种蛋白质的结合具有某种作为塞子的能力。它能阻止外排， 抗生素更有效。我们还创建了外膜桶的数据库，以更好地了解 这些蛋白质的序列结构关系。在这里，我们建议利用这些过去的成功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E.大肠杆菌抗生素外排泵，并确定确定的功能， 3）确定什么会促进蛋白质穿过外排泵的易位， 否则与外排泵结合。这项工作的全球概念，创造新的插头和易位， 膜β-桶，据我们所知，以前从未做过。这些活动的预期成果 实验是关于蛋白质-蛋白质相互作用如何促进堵塞和拉动的知识。 抗生素外排泵，以及通过TolC插入或拉动的肽的产生。这项工作是蓄势待发 因为它将证明重要的蛋白质-蛋白质相互作用 这是产生膜蛋白塞和转位器所必需的，从而可以使现有的 抗生素对抗抗药性超级细菌

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.