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Targeted TERS Investigations of Ligand-Receptor Binding

配体-受体结合的靶向 TERS 研究

基本信息

批准号：
9211336
负责人：
Zachary Schultz
金额：
$ 34.2万
依托单位：
UNIVERSITY OF NOTRE DAME
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 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9211336
关键词：
Accelerometer Address Adhesions Affinity Binding Biochemistry Cell membrane Cells Cellular Membrane Chemicals Comb animal structure Complex Coupling Data Detection Diffusion Disease Drug Targeting Environment Goals Gold Heterogeneity Immobilization In Situ In Vitro Individual Integrins Investigation Ligand Binding Ligands Location Measurement Membrane Membrane Proteins Metals Methodology Methods Molecular Monitor Nanostructures Peptides Pharmaceutical Preparations Pharmacologic Substance Property Proteins Receptor Signaling Reporting Research Resolution Role Signal Pathway Signal Transduction Specificity Spectrum Analysis Structure Surface Surface Plasmon Resonance System Techniques Technology Time Variant base experimental study imaging study improved insight instrumentation interest nanoparticle nanoscale new technology novel strategies particle plasmonics public health relevance receptor receptor binding spectroscopic survey tool trafficking

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to identify molecular interactions relevant to drug targeting and chemical signaling in cell membranes. More than half the drugs on the market target membrane proteins, a class of molecules where obtaining molecular detail in intact membranes challenges existing methods. We will use the plasmonic properties of nanoparticles to enable chemical-specific spectroscopic studies of ligand-receptor binding in intact cellular membranes. These investigations will demonstrate a new approach to probe the receptor's chemical residues that bind peptide antagonists and provide insight into molecular interactions that regulate the membrane proteins involved in signaling and drug targeting. Our approach combines tip enhanced Raman scattering (TERS), surface plasmon resonance (SPR) spectroscopy, and single particle tracking to characterize chemical interactions that regulate binding to membrane receptors. TERS will determine the chemical residues present in cell membranes that are associated with ligand binding. Initial results obtained in our lab indicate that ligand-functionalized nanoparticles selectively enhance the spectroscopic signals of the receptor involved in ligand binding. The ligand affinity can be determined from proteins immobilized on surfaces using SPR measurements. Complementary measurements can be performed in cells using single particle tracking. By taking advantage of technology that enables tracking of nanoparticles in real-time, we will characterize both ligand binding and membrane organization of individual ligand-functionalized nanoparticles bound to receptors on living cells. The specific aims of this proposal are: 1) Correlate in situ studies ligand affinity with the molecular identity of the receptor associated with peptide binding to the integrin receptors by combing TERS and SPR. 2) Explore protein receptor-binding interactions in vitro between integrin receptors and peptide ligands using targeted TERS. 3) Correlate in vitro binding affinity with chemical interaction by combining single nanoparticle tracking studies with TERS studies. The technology and platform we propose will address the challenge of obtaining chemical information from ligands binding to receptor proteins in intact cell membranes. 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.

描述（由申请人提供）：该提案的目标是确定与细胞膜中的药物靶向和化学信号传导相关的分子相互作用。市场上超过一半的药物以膜蛋白为目标，膜蛋白是一类分子，在完整膜中获取分子细节对现有方法提出了挑战。我们将利用纳米颗粒的等离子体特性来对完整细胞膜中的配体-受体结合进行化学特异性光谱研究。这些研究将展示一种新方法来探测与肽拮抗剂结合的受体化学残基，并深入了解调节参与信号传导和药物靶向的膜蛋白的分子相互作用。我们的方法结合了尖端增强拉曼散射（TERS）、表面等离子共振（SPR）光谱和单粒子追踪来表征调节与膜受体结合的化学相互作用。 TERS 将确定细胞膜中与配体结合相关的化学残基。我们实验室获得的初步结果表明，配体功能化纳米颗粒选择性增强参与配体结合的受体的光谱信号。可以使用 SPR 测量从固定在表面上的蛋白质确定配体亲和力。可以使用单粒子跟踪在细胞中进行补充测量。通过利用能够实时跟踪纳米颗粒的技术，我们将表征与活细胞上的受体结合的单个配体功能化纳米颗粒的配体结合和膜组织。该提案的具体目标是：1) 将原位研究配体亲和力与与肽结合到受体相关的受体的分子特性相关联。通过结合 TERS 和 SPR 整合素受体。 2) 使用靶向 TERS 探索整合素受体和肽配体之间的体外蛋白质受体结合相互作用。 3) 通过将单纳米粒子追踪研究与 TERS 研究相结合，将体外结合亲和力与化学相互作用相关联。我们提出的技术和平台将解决从与完整细胞膜中受体蛋白结合的配体获取化学信息的挑战。这些研究将为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相关提供新的见解。