Engineered Nanopores for Single-Molecule Stochastic Sensing

用于单分子随机传感的工程纳米孔

基本信息

  • 批准号:
    8760824
  • 负责人:
  • 金额:
    $ 28.64万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2009
  • 资助国家:
    美国
  • 起止时间:
    2009-09-28 至 2018-07-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Unraveling the interaction networks among functional proteins is essential in fundamental and clinical biomedical diagnostics by providing a mechanistic understanding of the complex regulatory processes of the cell, identifying their relationships to diseases, accelerating protein biomarker discovery, and assisting drug design. Advances in rational membrane protein design, chemical modification, biomolecular recognition, and single-molecule science will be used in concert for the creation of a new methodology to sample protein-protein interactions at high temporal and spatial resolution, as well as for the detection, exploration, and characterization of individual proteins. These proposed studies are aimed at engineering protein nanopore- based sensing devices featured by ligand-containing flexible tethers. Ample redesign of ferric hydroxamate uptake component A (FhuA), a monomeric b-barrel protein with a remarkable array of advantageous traits, such as robustness, versatility, and tractability, will result in a unique nanostructure with a single tethered proteinor DNA aptamer ligand at a strategic positioning of the nanopore. The FhuA-based scaffold is an attractive choice for this task, because it's open-state, quiet current remains stable for long periods within an unusually broad range of detection circumstances. These benefits will be used in various biosensing schemes, in which individual protein-protein and protein-DNA recognition events will produce detectable, discrete and reversible changes in the conformational dynamics of the movable tether, inducing alterations in the single- channel electrical signature. The expected immediate outcomes will be the following: (i) the creation of sensing elements for examining protein-protein interactions under equilibrium and non-equilibrium conditions; (ii) the development of highly specific nanopore-based sensing elements for a protein biomarker; (iii) a better understanding of the impact of tunable and constraining tethers on the intermolecular forces among protein partners, which has implications for the in vivo contexts of complex recognition events produced by anchored protein domains; (iv) the improvement in the sensitivity of the single-molecule detection of protein-protein interfaces, pushing forward the nanopore technology for the disentanglement of weak protein-protein interactions; (v) the expansion of the modularity and scalability of engineered protein nanopores as well as their integration with a synthetic membrane, improving their mechanical, thermal, electrical, and chemical stability. The adaptation of these unusual nanostructures with movable arms to an integrated microfabricated chip platform will provide a new generation of research tools for exploring the molecular basis of protein-protein recognition events in a sensitive, specific and quantitative fashion.
描述(由申请人提供):解开功能蛋白质之间的相互作用网络在基础和临床生物医学诊断中至关重要,因为它提供了对细胞复杂调控过程的机械理解,确定了它们与疾病的关系,加速了蛋白质生物标记物的发现,并帮助药物设计。合理的膜蛋白质设计、化学修饰、生物分子识别和单分子科学的进展将被用于创建一种新的方法学,以高时间和空间分辨率采样蛋白质-蛋白质相互作用,以及检测、探索和表征单个蛋白质。这些拟议的研究旨在设计基于蛋白质纳米孔的传感装置,其特点是含有配体的柔性系链。羟甲酸铁摄取成分A(FhuA)是一种单体b桶蛋白,具有一系列显著的优势特性,如坚固性、多功能性和易操纵性,对其进行充分的重新设计,将产生一种独特的纳米结构,在纳米孔的战略位置上具有单一的拴系蛋白或DNA适体配体。对于这项任务,基于FhuA的支架是一个有吸引力的选择,因为它的开态、安静的电流在异常广泛的检测环境中保持长时间稳定。这些优点将被用于各种生物传感方案,在这些方案中,单个蛋白质-蛋白质和蛋白质-DNA识别事件将在可移动系绳的构象动力学中产生可检测的、离散的和可逆的变化,导致单通道电信号的改变。预期的即时结果将如下:(I)创建用于检查平衡和非平衡条件下的蛋白质-蛋白质相互作用的传感元件;(Ii)开发用于蛋白质生物标记物的高度特定的基于纳米孔的传感元件;(Iii)更好地理解可调节和约束的系链对蛋白质伙伴之间的分子间力的影响,这对锚定的蛋白质结构域产生的复杂识别事件的体内环境具有影响;(Iv)提高蛋白质-蛋白质界面的单分子检测的灵敏度,推动纳米孔技术用于解开弱的蛋白质-蛋白质相互作用的纠缠;(V)扩大了工程蛋白纳米孔的模块化和可扩展性,以及它们与合成膜的集成,提高了它们的机械、热、电和化学稳定性。将这些具有可移动手臂的不同寻常的纳米结构改造成集成的微制造芯片平台,将为以灵敏、特定和定量的方式探索蛋白质-蛋白质识别事件的分子基础提供新一代研究工具。

项目成果

期刊论文数量(0)
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会议论文数量(0)
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LIVIU MOVILEANU其他文献

LIVIU MOVILEANU的其他文献

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{{ truncateString('LIVIU MOVILEANU', 18)}}的其他基金

Generalizable Nanosensors for Probing Highly Specific Interactions of Protein Kinases
用于探测蛋白激酶高度特异性相互作用的通用纳米传感器
  • 批准号:
    10719635
  • 财政年份:
    2023
  • 资助金额:
    $ 28.64万
  • 项目类别:
Development of Modular Synthetic Sensors for Protein Biomarker Detection
用于蛋白质生物标志物检测的模块化合成传感器的开发
  • 批准号:
    10659642
  • 财政年份:
    2023
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    10461887
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    7939932
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    8136461
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    8325070
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    8537210
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
Engineered Nanopores for Single-Molecule Stochastic Sensing
用于单分子随机传感的工程纳米孔
  • 批准号:
    10227053
  • 财政年份:
    2009
  • 资助金额:
    $ 28.64万
  • 项目类别:
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