Investigating G Protein Coupling of Nanodisc-solubilized Beta Adrenergic Receptor

研究纳米圆盘溶解的 β 肾上腺素受体的 G 蛋白偶联

基本信息

批准号：
8316845
负责人：
Courtney Sloan
金额：
$ 4.78万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-16 至 2013-05-02
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8316845
关键词：
Accounting Adipose tissue Adrenergic Receptor Agonist Annexins Arrestins Binding Binding Proteins Biological Biological Assay Biological Markers Biological Models Biological Process Blood Vessels Cardiac Coupling DNA Detection Detergents Development Devices Dimerization Drug Delivery Systems Drug Receptors Encapsulated Endocytosis Environment Event Family G-Protein-Coupled Receptors GTP-Binding Proteins Immobilization Ions Label Ligand Binding Ligands Lipid Bilayers Lipids Liver Malignant Neoplasms Marketing Mediating Membrane Membrane Proteins Methodology Methods Molecular Monitor Muscle Nutrient Optics Peptides Pharmacologic Substance Phospholipids Play Post-Translational Protein Processing Process Protein Binding Proteins R7 Virus Recombinants Refractive Indices Role Sampling Screening procedure Semiconductors Serine Signal Transduction Silicon Solutions Surface Technology Therapeutic Tissues Transducers Work base beta-adrenergic receptor drug development high throughput screening in vivo monoamine nanodisk nanoscale neurotransmission photonics prevent protein expression protein protein interaction receptor receptor binding receptor function response sensor therapeutic target

项目摘要

DESCRIPTION (provided by applicant): G protein coupled receptors (GPCRs) are ubiquitous signal transducers, highly sought after as pharmaceutical targets. Currently, half of all marketed therapeutics target GPCRs, and in many cases the off-target effects of these pharmaceutical products are also mediated through GPCRs. Due to their inherent instability and insolubility GPCRs are difficult to study; however, their essential role in biological processes certainly establishes this as a compelling and worthwhile pursuit. Lipid bilayer nanodiscs have emerged as a convenient format by which to solubilize membrane-bound proteins, mimicking their native environment while simultaneously stabilizing these molecules. Solubilizing GPCRs within the nanodisc construct necessitates the development of appropriate analytical assays capable of screening the ligand binding of multiple receptors and receptor subtypes. Silicon photonic devices such as optical microring resonators are highly multiplexable devices that are sensitive to small changes in refractive index near the ring surface. Biomolecules can be covalently attached to microrings and subsequent binding events can be characterized by monitoring each of the individually addressable microrings. This methodology offers the additional advantage of small sample volume requirements. By immobilizing nanodisc-solubilized ¿ adrenergic receptors onto the microring resonator platform, we aim to investigate G protein coupling and ¿ arrestin binding of these receptors in response to both agonists and antagonists. The effects of receptor dimerization will also be examined. By combining the nanodisc and microring resonator technologies, bioassays will be developed to characterize the effects of various ligands on GPCR binding and off-target effects in a highly multiplexable platform. PUBLIC HEALTH RELEVANCE: Approximately half of currently marketed pharmaceuticals target G protein coupled receptors (GPCRs). There is a significant need for highly multiplexed analysis platforms to characterize G protein coupling and arrestin binding of these receptors as indicators of potential off-target drug effects while maintaining the receptors in a native-like liid bilayer environment. In the proposed work, nanodisc-solubilized adrenergic receptors will be immobilized on silicon optical microring resonator platforms to investigate receptor binding in response to various ligands.

描述（由申请人提供）：G 蛋白偶联受体 (GPCR) 是普遍存在的信号转导器，作为药物靶点备受追捧。目前，一半的市售治疗药物都以 GPCR 为目标，并且在许多情况下，这些药品的脱靶效应是严重的。由于其固有的不稳定性和不溶性，GPCR 也通过 GPCR 介导；然而，它们在生物过程中的重要作用无疑使这一点成为一个令人信服且值得研究的课题。纳米圆盘已成为溶解膜结合蛋白的便捷形式，模拟其天然环境，同时稳定纳米圆盘结构内的 GPCR，因此需要开发能够筛选多种受体和受体的配体结合的适当分析方法。硅光子器件（例如光学微环谐振器）是高度可复用的器件，对环表面附近的折射率的微小变化敏感，可以是共价的。附着到微环和随后的结合事件可以通过监测每个可单独寻址的微环来表征，该方法通过固定纳米盘溶解的样品量提供了额外的优势。肾上腺素能受体到微环谐振器平台上，我们的目标是研究 G 蛋白耦合和 ¿通过结合纳米盘和微环谐振器技术，还将研究受体二聚化的影响，以表征各种配体对 GPCR 结合和脱靶的影响。高度可复用的平台中的效果。公共健康相关性：目前上市的药物中约有一半以 G 蛋白偶联受体 (GPCR) 为目标，因此迫切需要高度多重分析平台来表征这些受体的 G 蛋白偶联和抑制蛋白结合，作为潜在脱靶药物效应的指标。将受体维持在类似天然的液体双层环境中。在拟议的工作中，纳米盘溶解的肾上腺素能受体将被固定在硅光学微环上。谐振器平台用于研究响应各种配体的受体结合。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Interfacing lipid bilayer nanodiscs and silicon photonic sensor arrays for multiplexed protein-lipid and protein-membrane protein interaction screening.

DOI：
10.1021/ac3037359
发表时间：
2013-03-05
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Sloan, Courtney D. Kuhnline;Marty, Michael T.;Sligar, Stephen G.;Bailey, Ryan C.
通讯作者：
Bailey, Ryan C.

Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators.

DOI：
10.1021/ac300478f
发表时间：
2012-07-03
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Marty, Michael T.;Sloan, Courtney D. Kuhnline;Bailey, Ryan C.;Sligar, Stephen G.
通讯作者：
Sligar, Stephen G.