Enhanced Raman Imaging of Ligand-Receptor Recognition

配体受体识别的增强拉曼成像

基本信息

批准号：
10491043
负责人：
Zachary Schultz
金额：
$ 33.39万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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/10491043
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 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

项目摘要

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.

增强配体受体识别的拉曼成像抽象的：该建议的目的是确定与药物靶向和化学相关的分子相互作用活细胞中的信号传导。药物开发中的关键瓶颈是鉴定脱靶效应。可以识别与蛋白质相关的分子相互作用的方法蜂窝和其他现场模型中的候选人可用于理解和最大程度地减少不必要的副作用和并发症。在较早的药物筛查阶段确定这些影响对于避免后期很重要药物衰竭。我们正在开发利用金属塑料特性的技术纳米颗粒能够对活细胞中配体受体结合的化学特异性光谱研究。这些调查将提供新的方法来探测接收器的化学保留率，以结合肽拮抗剂，并提供有关调节信号传导涉及的蛋白质的分子相互作用的见解药物靶向。我们的方法结合了两种纳米颗粒的增强拉曼散射（表面增强的拉曼）散射，或SERS）和扫描问题（尖端增强的拉曼散射或TER），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级分辨率SERS成像以表征调节与蛋白质受体结合的化学相互作用。在增强的拉曼分散响应中存在的信息提供了分子水平的细节蛋白质识别识别的相互作用。在共同的情况下，我们的方法提供了一种新的方法监测蛋白质与活细胞中假定药物的结合，并表征靶向特异性。该提案的具体目的是： 1）筛选活细胞中胡椒官能化纳米颗粒的靶向特异性。 2）开发超分辨率的SERS成像以改善结合特异性研究并确定细胞中的粒子位置。 3）将非标准量子和数值模拟与实验相结合以识别钥匙与肽结合有关的氨基酸构象基序。总体而言，我们提出的技术和平台将应对获取化学信息的挑战从完整细胞中的配体与受体蛋白结合。这些研究将为您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如何与疾病和治疗有关。