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（由适用提供）：该提案的目的是确定与细胞膜中药物靶向和化学信号相关的分子相互作用。市场上的一半以上药物靶向膜蛋白，这是一类分子，其中获得完整膜中分子细节的分子挑战了现有方法。我们将使用纳米颗粒的等离子体特性来实现完整细胞膜中配体受体结合的化学特异性光谱研究。这些研究将证明一种新方法来探测受体的化学保留率，该化学保留量结合肽拮抗剂，并提供对调节信号传导和药物靶向涉及的膜蛋白的分子相互作用的洞察力。我们的方法结合了尖端增强的拉曼散射（TER），表面等离子体共振（SPR）光谱和单个颗粒跟踪，以表征调节与膜受体结合的化学相互作用。 TER将确定与配体结合相关的细胞膜中存在的化学结果。在我们的实验室中获得的初始结果表明，配体官能化纳米颗粒有选择地增强了参与配体结合的受体的光谱信号。配体亲和力可以通过使用SPR测量在表面上固定在表面上的蛋白质确定。可以使用单个粒子跟踪在细胞中进行互补测量。通过利用能够实时跟踪纳米颗粒的技术，我们将表征配体结合和膜组织的单个配体功能化纳米颗粒与活细胞上受体结合的纳米颗粒。该提案的具体目的是：1）与原位研究配体亲和力与与肽结合与与肽结合相关的受体的分子身份相关通过组合TER和SPR组成蛋白受体。 2）使用靶向TERS探索整联蛋白受体和肽配体之间的蛋白质受体结合相互作用。 3）通过将单纳米颗粒跟踪研究与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完整细胞膜中与受体蛋白结合的配体结合的化学信息的挑战。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与疾病和治疗有关。