Chemical probes to modulate acyl-homoserine lactone quorum signal synthesis

调节酰基高丝氨酸内酯群体信号合成的化学探针

• 批准号: 10579520
• 负责人: Rajesh Nagarajan
• 金额: $ 40.86万
• 依托单位: BOISE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579520
• 关键词: Academic Research Enhancement Awards; Active Sites; Acyl Carrier Protein; Acyl Coenzyme A; Allosteric Site; Applications Grants; Bacteria; Binding; Biological Assay; Biomedical Research; Cell Communication; Cells; Chemicals; Coenzyme A; Coupling; Data; Diffuse; Docking; Environment; Enzymes; Exposure to; Foundations; Funding; Future; Goals; Gram-Negative Bacteria; Grant; High Pressure Liquid Chromatography; Home; Homocysteine; In Vitro; Institution; Interruption; Kinetics; Libraries; Maps; Mediating; Molecular; National Institute of General Medical Sciences; Outcome; Pathogenicity; Physicians; Pilot Projects; Population Density; Proteins; Pseudomonas aeruginosa; Research; Research Infrastructure; S-Adenosylmethionine; Signal Transduction; Specificity; Substrate Specificity; Testing; Training; United States National Institutes of Health; Virulence; analog; biochemical tools; career; design; homoserine lactone; in vivo; infection rate; inhibitor; multi-drug resistant pathogen; novel; programs; quorum sensing; rational design; receptor; small molecule; small molecule inhibitor; tool; undergraduate student

Abstract Gram-negative bacteria count specific, N-acyl-L-homoserine lactone (AHL) quorum sensing (QS) signals in the environment to estimate local population densities, facilitate cell-cell communication and virulence. AHL synthases make AHL autoinducer signals by enzymatically coupling acyl-ACP/acyl-CoA and S-adenosyl-L- methionine metabolites to facilitate quorum sensing for the bacterium. Small molecule inhibitors of AHL synthase enzymes would limit signal synthesis, interrupt quorum sensing and thus hold significant promise as chemical tools to investigate QS networks in bacteria. Designing AHL synthase-specific inhibitors, however, does remain a significant challenge. To develop AHL synthase selective inhibitors, we recently embarked on a proof-of-concept study to evaluate the potential of AHL signal derivatives as product analog inhibitors of Pseudomonas aeruginosa RhlI AHL synthase. In this study, we demonstrated that AHL analogs could serve as ‘chemical probes’ to uncover novel inhibition and activation binding pockets that could be tapped to develop potent AHL synthase-specific modulators. In this grant application, we extend this approach on a broader scale to evaluate the utility of AHL analog chemical probes in discovering novel binding pockets, determining the mechanistic basis of AHL synthase modulation, and formulating the rules of AHL engagement in short, medium and long-chain preferring AHL synthases. Additionally, we will determine how the inhibition and activation pockets could be leveraged to develop novel substrates and substrate analog inhibitors of AHL synthases. Successful completion of these studies should inform the rational design of inhibition and activation pocket directed AHL synthase modulators. Finally, the goals described in this application are tightly aligned with the objectives of AREA program, which are to a) support meritorious research at an undergraduate focused institution, b) strengthen the research environment at the home institution and c) provide opportunity for undergraduate students to get involved in biomedical research.

摘要 革兰氏阴性菌计数特异性，N-酰基-L-高丝氨酸内酯（阿勒）群体感应（QS）信号在 环境，以估计当地人口密度，促进细胞间的沟通和毒力。阿勒 腺苷酸酶通过酶促偶联酰基-ACP/酰基-CoA和S-腺苷-L-腺苷酸， 甲硫氨酸代谢物，以促进细菌的群体感应。阿勒的小分子抑制剂 合成酶将限制信号合成，中断群体感应，因此具有重要的前景 作为研究细菌QS网络的化学工具。然而，设计阿勒合酶特异性抑制剂， 仍然是一个巨大的挑战。为了开发阿勒合酶选择性抑制剂，我们最近开始 一项概念验证研究，旨在评估阿勒信号衍生物作为产品类似物抑制剂的潜力， 铜绿假单胞菌RhII阿勒合酶。在这项研究中，我们证明了阿勒类似物可以作为 作为“化学探针”，以发现新的抑制和激活结合口袋，可以开发 有效的阿勒转氨酶特异性调节剂。在这项拨款申请中，我们将这种方法扩展到更广泛的领域。 评估阿勒类似物化学探针在发现新结合口袋中的效用的量表， 阿勒合成酶调节的机制基础，并制定阿勒参与的规则，简而言之， 中链和长链偏好阿勒酶。此外，我们将确定如何抑制和 激活口袋可以用来开发新的底物和底物类似物阿勒抑制剂 - 是的这些研究的成功完成应该为抑制和激活的合理设计提供信息 口袋定向阿勒合酶调节剂。最后，本应用程序中描述的目标是紧密一致的 与区域计划的目标，这是a）支持在本科有价值的研究 重点机构，B）加强国内机构的研究环境和c）提供机会 让本科生参与生物医学研究。