Enhanced raman imaging of ligand-receptor recognition

配体-受体识别的增强拉曼成像

• 批准号: 10596420
• 负责人: Zachary Schultz
• 金额: $ 13.21万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596420
• 关键词: Address; Adsorption; Affinity; Amino Acids; Binding; Binding Proteins; Biochemistry; Biological; Biological Assay; Cells; Chemicals; Complement; Complex; Detection; Disease; Drug Screening; Drug Targeting; Environment; Failure; Fluorescence Microscopy; Gene Mutation; Goals; Gold; Human; Image; Imaging ligands; Investigation; Ligand Binding; Ligands; Location; Metals; Methodology; Methods; Microscopy; Modeling; Molecular; Molecular Conformation; Monitor; Organism; Outcome; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Property; Proteins; Public Health; Raman Spectrum Analysis; Research; Resolution; Scanning; Serum Proteins; Signal Pathway; Signal Transduction; Specificity; Surface; System; Techniques; Technology; Therapeutic; antagonist; base; biological systems; drug candidate; drug development; drug efficacy; experimental study; improved; insight; instrumentation; metallicity; molecular imaging; nanoparticle; nanoscale; new technology; novel strategies; particle; plasmonics; quantum; receptor; receptor binding; response; side effect; simulation; spectroscopic survey; tool

Enhanced Raman Imaging of Ligand-Receptor Recognition Abstract: The goal of this proposal is to identify molecular interactions relevant to drug targeting and chemical signaling in living cells. A critical bottleneck in the development of drugs is the identification of off-target effects. Methods that can identify the molecular interactions associated with proteins recognizing and binding to drug candidates in cellular and other live models can be used to understand and minimize unwanted side effects and complications. Identifying these effects at earlier stages of drug screening is important to avoid late stage drug failure. We are developing technologies that take advantage of the plasmonic properties of metallic nanoparticles to enable chemical-specific spectroscopic studies of ligand-receptor binding in living cells. These investigations will provide new approaches to probing the receptor’s chemical residues that bind peptide antagonists and provide insights into molecular interactions that regulate the proteins involved in signaling and drug targeting. Our approach combines enhanced Raman scattering from both nanoparticles (Surface enhanced Raman scattering, or SERS) and scanning probes (tip enhanced Raman scattering, or TERS), nanoparticle tracking microscopy, non-standard applications of static and dynamic quantum chemical calculations, and super- resolution SERS imaging to characterize chemical interactions that regulate binding to protein receptors. Information present in the enhanced Raman scattered response provides molecular level detail of the interactions governing recognition by the protein. In concert, our methodologies provide a new approach to monitoring protein binding to putative drugs in living cells, and to characterize targeting specificity. The specific aims of this proposal are: 1) Screen the targeting specificity of peptide-functionalized nanoparticles in live cells. 2) Develop super-resolution SERS imaging to improve binding specificity studies and identify particle location in cells. 3) Combine non-standard quantum and numerical simulations with experiments to identify key motifs in amino acid conformation related to peptide binding. Overall, the technology and platform we propose will address the challenge of obtaining chemical information from ligands binding to receptor proteins in intact cells. These studies will provide new insights into the molecular interactions that regulate signaling pathways and how anomalies in these interactions are associated with disease and treatment.

配体-受体识别的增强拉曼成像 摘要： 该提案的目标是确定与药物靶向和化学药物靶向相关的分子相互作用。 活细胞中的信号。药物开发的一个关键瓶颈是脱靶效应的识别。 能够鉴定与蛋白质识别和结合药物相关的分子相互作用的方法 细胞模型和其他实时模型中的候选者可用于了解和最小化不必要的副作用 和并发症。在药物筛选的早期阶段确定这些影响对于避免晚期阶段非常重要。 药物失效我们正在开发利用金属等离子体特性的技术， 纳米颗粒，使化学特异性光谱研究的配体受体结合在活细胞中。这些 研究将提供新的方法来探测受体结合肽的化学残基 拮抗剂，并提供了对分子相互作用的见解，调节参与信号传导的蛋白质， 药物靶向 我们的方法结合了来自两种纳米颗粒的增强拉曼散射（表面增强拉曼散射）。 散射，或Sers）和扫描探针（尖端增强拉曼散射，或TERS），纳米颗粒跟踪 显微镜，静态和动态量子化学计算的非标准应用，以及超 分辨率Sers成像，以表征调节与蛋白质受体结合的化学相互作用。 存在于增强的拉曼散射响应中的信息提供了该拉曼光谱的分子水平细节。 控制蛋白质识别的相互作用。与此同时，我们的方法提供了一种新的方法， 监测活细胞中蛋白质与推定药物的结合，并表征靶向特异性。 这项建议的具体目标是： 1)筛选肽功能化纳米粒子在活细胞中的靶向性。 2)开发超分辨率Sers成像，以改善结合特异性研究并识别 细胞中的粒子位置。 3)联合收割机非标准量子和数值模拟与实验相结合， 与肽结合相关的氨基酸构象中的基序。 总的来说，我们提出的技术和平台将解决获得化学信息的挑战 从配体结合到完整细胞中的受体蛋白。这些研究将提供新的见解， 调节信号通路的分子相互作用，以及这些相互作用中的异常是如何 与疾病和治疗有关。