Discovery of New Antibacterial Targets: Probes and Inhibitors of Histidine Kinase Proteins

细菌依靠信号转导途径对环境信号做出反应。他们有能力具体地 并灵敏地检测背景中单个化学实体的存在或不存在 数以千计的其他分子对它们的生存至关重要。双组分系统(TCS；通常为20-120 每个有机体的离散系统)对于将这种复杂的分子环境转化为 细菌的作用，从生长到抗生素耐药性再到毒性。尽管TCSs对 生物体的适应性反应，技术挑战阻碍了它们的研究，限制了我们对 小区如何协调来自多个TC的信号的组合传输以协调 生存和发病机制。 少数TCS已被确定为开发抗菌药物的重要靶点 探员们。然而，很明显，细菌信号的复杂过程包含许多额外的靶点 将是与迅速到来的“后抗生素时代”作斗争的关键。拟议研究的目标是 开发和应用研究组氨酸激酶调节所需的工具和技术，关键是 TCS介导的细菌信号转导所需的蛋白质。组氨酸激酶直接感知外部刺激和 在细胞内传递这种化学信息，促进基因表达的调节。归根结底， 在这项工作中积累的工具和知识将使我们能够理解和预测信号是如何传播的 转化为细菌活动，并劫持TCSs用于治疗传染病。这些目标将是 通过追求三个目标来实现。目标1.开发基于ATP的分子作为基于活性的探针 组氨酸激酶活性的评估。目的2.从环境中全球定位组氨酸激酶的激活 应激源、抗生素暴露和宿主相互作用，以确定快速细菌所需的关键HKS 适应和生存。目的3.利用多组氨酸激酶抑制剂来确定主要的蛋白质 致病性和毒力表型的贡献者。 显然，迫切需要新的方法来治疗抗生素耐药感染- 抗生素流水线将重点放在“死亡表型”上，这导致了抗生素的迅速崛起 抗药性和严重缺乏通过新的作用机制发挥作用的分子。控制 行为表型，尤其是致病机理和致病力，是一个非常有希望但尚未实现的研究方向。 控制传染病的方法。我们高度跨学科和创新的方法将使 美国将汇集对TCS介导的信号转导的批判性理解，并特别强调 致病和毒力相关过程中所需的HKS的特征。

Bacteria rely on signal transduction pathways to respond to environmental cues. Their ability to specifically and sensitively detect the presence or absence of a single chemical entity amongst the background of thousands of other molecules is essential for their survival. Two-component systems (TCSs; generally 20-120 discrete systems per organism) are crucial for the translation of this complex molecular environment into bacterial action, ranging from growth to antibiotic resistance to virulence. Despite the central role of TCSs to an organism’s adaptive response, technical challenges have hampered their study, limiting our understanding of how the combined transmission of the signals from many TCSs are coordinated by the cell to orchestrate survival and pathogenesis. A small number of TCSs have been identified as important targets for the development of antibacterial agents. However, it is clear that the complex process of bacterial signaling holds many additional targets that will be critical for combating the rapidly approaching “post-antibiotic era.” The goal of the proposed research is to develop and apply the tools and techniques required to study the regulation of the histidine kinases, the key proteins required for TCS-mediated bacterial signaling. Histidine kinases directly sense external stimuli and transmit this chemical message inside of the cell, promoting the regulation of gene expression. Ultimately, the tools and knowledge amassed in this work will enable us to understand and predict how a signal is propagated into bacterial action and to hijack the TCSs for the treatment of infectious disease. These goals will be accomplished by pursuit of three Aims. Aim 1. Develop ATP-based molecules as activity-based probes for assessment of histidine kinase activation. Aim 2. Globally map histidine kinase activation from environmental stressors, antibiotic exposure, and host interactions to identify the key HKs required for rapid bacterial adaptation and survival. Aim 3. Utilize multi-histidine kinase inhibitors to determine the proteins that are major contributors to pathogenicity and virulence phenotypes. It is clear that there is a dire need for new approaches to the treatment of antibiotic-resistant infections – the antibiotic pipeline has focused on the “death phenotype,” which has led to the rapid rise of antibiotic resistance and a severe lack of molecules that function through novel mechanisms-of-action. Control of behavioral phenotypes, especially pathogenesis and virulence, is an exceptionally promising but unrealized approach for the control of infectious disease. Our highly interdisciplinary and innovative approach will enable us to assemble a critical understanding of TCS-mediated signal transduction, with particular emphasis on characterization of the HKs required in pathogenesis- and virulence-associated processes.