Advancing ribosome-targeting antibacterial peptides with a unique mechanism of action

以独特的作用机制推进核糖体靶向抗菌肽

基本信息

  • 批准号:
    10569098
  • 负责人:
  • 金额:
    $ 75万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2022
  • 资助国家:
    美国
  • 起止时间:
    2022-02-08 至 2027-01-31
  • 项目状态:
    未结题

项目摘要

Project Summary Apidaecin (Api) and Drosocin (Dro), are proline-rich antimicrobial peptides (PrAMPs) produced by honeybees and fruit flies, respectively, which share a unique mechanism of action. Our previous studies of Api showed that upon entering Gram-negative bacterial cells through the SbmA transporter, Api binds in the exit tunnel of ribosomes that have just released the newly made protein and arrests the ribosomes at stop codons by trapping the associated tRNA and release factor. As such, Api represents the first-ever described specific inhibitor of translation termination. Our subsequent whole-genome studies revealed that arresting terminating ribosomes triggers several downstream events that accentuate the inhibitory action of this PrAMP, including ribosome queuing and readthrough of stop codons. Our preliminary data indicate that Dro, despite its distinct amino acid sequence, inhibits the termination step of translation as well, by a mechanism likely resembling that of Api. Their idiosyncratic mode of binding to the target, the unique mechanism of action, and the triggering of downstream effects harmful for the bacterial cell, make these antibacterial peptides an attractive model for developing novel antibiotics. Furthermore, the biological nature of these PrAMPs opens unique opportunities for their screening and optimization by generating hundreds of thousands of peptide variants directly in bacterial cells. In the current proposal we will use the combined effort of three laboratories with expertise in biochemistry and genomics of ribosomal antibiotics, in peptide chemistry and in structural analysis of ribosome-antibiotic complexes to advance the fundamental understanding of the mechanism of action of Api- and Dro-like translation termination inhibitors and identify derivatives with superior on-target activity and expanded spectrum of antibacterial action. In order to achieve these goals we will test arrays of Api and Dro variants in bacterial cells by the tunable expression of peptide gene libraries, determine high-resolution X-ray crystal structures of ribosome-peptide complexes, and employ rational structure-based design to generate via chemical synthesis peptide variants with superior properties. Specifically: In Aim 1, we will identify Api-derived peptides with improved activity upon ribosomes from Gram-negative and Gram-positive pathogens. In Aim 2, the spectrum of action of Api-like peptides will be expanded by bypassing the necessity for uptake by the SbmA transporter. Finally, in Aim 3, we will analyze the ribosome binding and mechanism of action of Dro-like peptides and use comparative analysis to identify the key features that define the class of antimicrobial peptides that target translation termination. The three Aims are tightly interconnected but completely independent from each other. The reagents and tools that will be generated in the course of the proposed work are aimed to serve as leads for future clinical development. Importantly, the results obtained in the proposed studies will significantly advance the fundamental understanding of the properties and mechanisms of action of PrAMPs and will stimulate the progress of the field of ribosome-targeting antibacterial peptides, which currently is still in its infancy.
项目摘要 Apidaecin(Api)和Droocin(Dro)是蜜蜂和蜜蜂产生的富含脯氨酸的抗菌肽(PRAMP)。 果蝇,它们都有一个独特的作用机制。我们之前对Api的研究表明,在进入 革兰氏阴性细菌细胞通过SBMA转运蛋白,Api结合在刚刚释放的核糖体的出口隧道中 这种新合成的蛋白质通过捕获相关的tRNA和释放因子,在终止密码子上阻止核糖体。AS 这样,Api代表了第一个被描述的翻译终止的特异性抑制物。我们随后的全基因组 研究表明,阻止终止核糖体触发了几个下游事件,从而加剧了抑制作用 PrAMP的作用,包括核糖体排队和停止密码子的通读。我们的初步数据显示, 尽管它有不同的氨基酸序列,但也抑制了翻译的终止步骤,其机制很可能类似于 那是阿比的。它们与靶标结合的独特模式,独特的作用机制,以及 下游效应对细菌细胞有害,使这些抗菌肽成为开发新型抗菌肽的诱人模型 抗生素。此外,这些野马的生物学特性为它们的筛选和 通过在细菌细胞中直接产生数十万个多肽变体进行优化。 在目前的提案中,我们将使用三个拥有生物化学和专业知识的实验室的联合努力 核糖体抗生素基因组学,在多肽化学和核糖体-抗生素复合体的结构分析中 加深对Api和Dro样翻译终止抑制剂作用机制的基本认识 并鉴定具有良好的靶向活性和广谱抗菌作用的衍生物。为了实现 这些目标我们将通过肽基因文库的可调表达在细菌细胞中测试Api和dro变体的阵列, 确定核糖体-多肽复合体的高分辨X射线晶体结构,并采用合理的结构基础 设计通过化学合成产生具有优越性能的多肽变体。具体来说:在目标1中,我们将确定 来自革兰氏阴性和革兰氏阳性病原体的API衍生多肽对核糖体的活性提高。在目标2中, Api样肽的作用范围将通过绕过SBMA转运体摄取的必要性而扩大。 最后,在目标3,我们将分析核糖体结合和作用机制,并使用比较 分析以确定定义以翻译终止为目标的抗菌肽类别的关键特征。这个 三个目标紧密相连,但又完全相互独立。 将在拟议工作过程中产生的试剂和工具旨在作为以下方面的线索 未来的临床发展。重要的是,拟议研究的结果将大大推进 从根本上了解PRMP的性质和作用机制,并将推动 核糖体靶向抗菌肽领域,目前还处于起步阶段。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

ALEXANDER S MANKIN其他文献

ALEXANDER S MANKIN的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('ALEXANDER S MANKIN', 18)}}的其他基金

Advancing ribosome-targeting antibacterial peptides with a unique mechanism of action
以独特的作用机制推进核糖体靶向抗菌肽
  • 批准号:
    10443921
  • 财政年份:
    2022
  • 资助金额:
    $ 75万
  • 项目类别:
Advancing ribosome-targeting antibacterial peptides with a unique mechanism of action
以独特的作用机制推进核糖体靶向抗菌肽
  • 批准号:
    10436039
  • 财政年份:
    2021
  • 资助金额:
    $ 75万
  • 项目类别:
Equipment Supplement Request for Purchasing Amersham Typhoon RGB Phosphorimager (for R35GM127134)
购买 Amersham Typhoon RGB 荧光成像仪(适用于 R35GM127134)的设备补充申请
  • 批准号:
    10386084
  • 财政年份:
    2018
  • 资助金额:
    $ 75万
  • 项目类别:
Exploiting antibiotics to understand the ribosome and translation
利用抗生素来了解核糖体和翻译
  • 批准号:
    10366000
  • 财政年份:
    2018
  • 资助金额:
    $ 75万
  • 项目类别:
Exploiting antibiotics to understand the ribosome and translation
利用抗生素来了解核糖体和翻译
  • 批准号:
    9897557
  • 财政年份:
    2018
  • 资助金额:
    $ 75万
  • 项目类别:
Context-specific action of antibiotics targeting the catalytic center of the bacterial ribosome
针对细菌核糖体催化中心的抗生素的特定作用
  • 批准号:
    9158354
  • 财政年份:
    2016
  • 资助金额:
    $ 75万
  • 项目类别:
Context-specific action of antibiotics targeting the catalytic center of the bacterial ribosome
针对细菌核糖体催化中心的抗生素的特定作用
  • 批准号:
    9332339
  • 财政年份:
    2016
  • 资助金额:
    $ 75万
  • 项目类别:
Molecular mechanisms of action of macrolide antibiotics
大环内酯类抗生素的分子作用机制
  • 批准号:
    8482422
  • 财政年份:
    2013
  • 资助金额:
    $ 75万
  • 项目类别:
Molecular mechanisms of action of macrolide antibiotics
大环内酯类抗生素的分子作用机制
  • 批准号:
    8640960
  • 财政年份:
    2013
  • 资助金额:
    $ 75万
  • 项目类别:
Programmed translation arrest controlled by nascent peptides and antibiotics
由新生肽和抗生素控制的程序化翻译停滞
  • 批准号:
    8917273
  • 财政年份:
    2012
  • 资助金额:
    $ 75万
  • 项目类别:

相似海外基金

RII Track-4:NSF: From the Ground Up to the Air Above Coastal Dunes: How Groundwater and Evaporation Affect the Mechanism of Wind Erosion
RII Track-4:NSF:从地面到沿海沙丘上方的空气:地下水和蒸发如何影响风蚀机制
  • 批准号:
    2327346
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Standard Grant
BRC-BIO: Establishing Astrangia poculata as a study system to understand how multi-partner symbiotic interactions affect pathogen response in cnidarians
BRC-BIO:建立 Astrangia poculata 作为研究系统,以了解多伙伴共生相互作用如何影响刺胞动物的病原体反应
  • 批准号:
    2312555
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Standard Grant
How Does Particle Material Properties Insoluble and Partially Soluble Affect Sensory Perception Of Fat based Products
不溶性和部分可溶的颗粒材料特性如何影响脂肪基产品的感官知觉
  • 批准号:
    BB/Z514391/1
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Training Grant
Graduating in Austerity: Do Welfare Cuts Affect the Career Path of University Students?
紧缩毕业:福利削减会影响大学生的职业道路吗?
  • 批准号:
    ES/Z502595/1
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Fellowship
Insecure lives and the policy disconnect: How multiple insecurities affect Levelling Up and what joined-up policy can do to help
不安全的生活和政策脱节:多种不安全因素如何影响升级以及联合政策可以提供哪些帮助
  • 批准号:
    ES/Z000149/1
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Research Grant
感性個人差指標 Affect-X の構築とビスポークAIサービスの基盤確立
建立个人敏感度指数 Affect-X 并为定制人工智能服务奠定基础
  • 批准号:
    23K24936
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
How does metal binding affect the function of proteins targeted by a devastating pathogen of cereal crops?
金属结合如何影响谷类作物毁灭性病原体靶向的蛋白质的功能?
  • 批准号:
    2901648
  • 财政年份:
    2024
  • 资助金额:
    $ 75万
  • 项目类别:
    Studentship
ERI: Developing a Trust-supporting Design Framework with Affect for Human-AI Collaboration
ERI:开发一个支持信任的设计框架,影响人类与人工智能的协作
  • 批准号:
    2301846
  • 财政年份:
    2023
  • 资助金额:
    $ 75万
  • 项目类别:
    Standard Grant
Investigating how double-negative T cells affect anti-leukemic and GvHD-inducing activities of conventional T cells
研究双阴性 T 细胞如何影响传统 T 细胞的抗白血病和 GvHD 诱导活性
  • 批准号:
    488039
  • 财政年份:
    2023
  • 资助金额:
    $ 75万
  • 项目类别:
    Operating Grants
How motor impairments due to neurodegenerative diseases affect masticatory movements
神经退行性疾病引起的运动障碍如何影响咀嚼运动
  • 批准号:
    23K16076
  • 财政年份:
    2023
  • 资助金额:
    $ 75万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了