UV Plasmon-Enhanced Chiroptical Spectroscopy of Membrane-Binding Proteins

膜结合蛋白的紫外等离子增强手性光谱

基本信息

批准号：
10680969
负责人：
Bjoern Markus Reinhard
金额：
$ 41.2万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-23 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10680969
关键词：
Aluminum Binding Binding Proteins Binding Sites Biological Biological Process Buffers Chemicals Circular Dichroism Circular Dichroism Spectroscopy Cryoelectron Microscopy Crystallization Disease Electromagnetic Fields Electromagnetics Electronics Engineering Environment Equilibrium Frequencies Goals Gold Higher Order Chromatin Structure Hot Spot Individual Investigation Lead Light Lipids Magnetism Measurement Membrane Membrane Fusion Membrane Lipids Membrane Proteins Methods Microscope Molecular Molecular Structure Nanostructures Nucleic Acids Optics Performance Peripheral Pharmacologic Substance Phase Physiological Preparation Property Proteins Public Health Raman Spectrum Analysis Research Sampling Signal Transduction Silver Spectrum Analysis Structure Surface Surface Plasmon Resonance Surface Properties Techniques Temperature Tertiary Protein Structure Testing Vibrational Circular Dichroism X ray spectroscopy X-Ray Crystallography absorption alpha synuclein amphiphilicity aqueous biophysical properties biophysical tools chiral molecule design disease diagnostic enantiomer improved insight instrument interest membrane activity miniaturize nano nanoengineering nanoparticle plasmonics protein structure prototype small molecule stereochemistry temporal measurement theories tool ultraviolet

Aluminum Binding Binding Proteins Binding Sites Biological Biological Process Buffers Chemicals Circular Dichroism Circular Dichroism Spectroscopy Cryoelectron Microscopy Crystallization Disease Electromagnetic Fields Electromagnetics Electronics Engineering Environment Equilibrium Frequencies Goals Gold Higher Order Chromatin Structure Hot Spot Individual Investigation Lead Light Lipids Magnetism Measurement Membrane Membrane Fusion Membrane Lipids Membrane Proteins Methods Microscope Molecular Molecular Structure Nanostructures Nucleic Acids Optics Performance Peripheral Pharmacologic Substance Phase Physiological Preparation Property Proteins Public Health Raman Spectrum Analysis Research Sampling Signal Transduction Silver Spectrum Analysis Structure Surface Surface Plasmon Resonance Surface Properties Techniques Temperature Tertiary Protein Structure Testing Vibrational Circular Dichroism X ray spectroscopy X-Ray Crystallography absorption alpha synuclein amphiphilicity aqueous biophysical properties biophysical tools chiral molecule design disease diagnostic enantiomer improved insight instrument interest membrane activity miniaturize nano nanoengineering nanoparticle plasmonics protein structure prototype small molecule stereochemistry temporal measurement theories tool ultraviolet

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项目摘要

Summary Circular dichroism (CD) and Raman optical activity (ROA) are chiroptical spectroscopies that provide valuable structural information about biomolecules and pharmaceuticals under native conditions in aqueous buffer without the need for special sample preparation or crystallization. The two methods are complementary as they probe the circular dichroism of molecular electronic and molecular vibrational transitions, respectively. A combination of the two methods is particularly well suited for investigating the structure of membrane binding proteins, which remain very difficult to characterize with other biophysical characterization tools. Although in theory a combined CD / ROA characterization has the potential for providing important structural information of membrane binding proteins, in practice the weak sensitivities of the two spectroscopies makes it difficult to realize this potential. A need for high sample concentrations and long acquisition times has limited a more widespread use of CD and in particular ROA spectroscopy as tool for characterizing membrane binding proteins. This project intends to overcome the sensitivity limitations of CD and ROA spectroscopies by developing plasmon-enhanced CD (PECD) and surface-enhanced ROA (SEROA) spectroscopies that utilize plasmonic nanoantennas, which are engineered nanostructures with specific electric (E) and magnetic (H) field properties as well as defined phase properties, to enhance signal intensities. To maximize the signal enhancement, antenna substrates will be developed with plasmon resonances in the ultraviolet (UV) so that the electromagnetic resonances can overlap with the molecular electronic resonances of biological target molecules, facilitating strong signal intensities for both CD and ROA. As this proposal focuses on developing PECD and SEROA as characterization tool for membrane binding proteins, another important design component of the proposed antennas is the assembly of a lipid membrane on the surface of the plasmonic nanoantennas to provide binding sites for membrane binding proteins. This approach enriches the proteins of interest in electromagnetic hot spots where CD and ROA signal enhancements are highest and allows for a spectroscopic characterization of the protein structure in its membrane-bound form. The developed plasmon-enhanced spectroscopies will enable important new insights into the structure and chirality of membrane-binding proteins, for instance as function of lipid compositions, and will contribute to a greatly improved understanding of protein-membrane interactions. The specific aims of this application are to: Aim 1: Develop a Plasmon-Enhanced Ultraviolet CD Spectroscopy for Membrane Binding Proteins Aim 2: Develop Plasmon Enhanced Raman Optical Activity (ROA) Spectroscopy for Membrane Binding Proteins Aim 3: Prototype Combined Electronic CD / ROA Instrument for the Characterization of Membrane Binding Proteins

概括圆二色性（CD）和拉曼光学活动（ROA）是手势光谱，可提供有价值的光谱在没有水性缓冲液中的天然条件下，有关生物分子和药品的结构信息需要特殊样品制备或结晶的需求。这两种方法是互补的分子电子和分子振动转变的圆形二色性。组合这两种方法特别适合研究膜结合蛋白的结构，使用其他生物物理表征工具来表征非常困难。虽然从理论上讲 CD / ROA表征有可能提供膜结合的重要结构信息蛋白质在实践中，两种光谱镜的敏感性较弱，因此很难意识到这一潜力。一个需要高样本浓度和长期获取时间的需求限制了CD的广泛使用和特别是ROA光谱作为表征膜结合蛋白的工具。这个项目打算通过开发等离子体增强CD来克服CD和ROA光谱的灵敏度限制（PECD）和表面增强的ROA（Seroa）光谱镜，利用等离子纳米antennas 具有特定电气（E）和磁场特性的工程纳米结构以及定义的相位属性，以增强信号强度。为了最大化信号增强，天线底物将是用紫外线（UV）中的等离子共振开发，以使电磁共振可以重叠随着生物靶标分子的分子电子共振，促进了强信号强度 CD和ROA。由于该建议着重于开发PECD和Seroa作为特征工具膜结合蛋白，所提出天线的另一个重要设计组成部分是组装等离激元纳米antennans表面上的脂质膜，以提供膜结合的结合位点蛋白质。这种方法丰富了电磁热点中感兴趣的蛋白质，其中CD和ROA信号增强功能是最高的，可以在其蛋白质结构中进行光谱表征膜结合形式。开发的等离子增强光谱镜将使重要的新见解进入膜结合蛋白的结构和手性，例如作为脂质组成的功能，而将有助于对蛋白质 - 膜相互作用的大大改善。这个特定的目的应用程序是： AIM 1：为膜结合蛋白开发等离子体增强的紫外线CD光谱 AIM 2：开发血浆增强的拉曼光活性（ROA）光谱膜的膜结合蛋白 AIM 3：原型组合电子CD / ROA仪器，用于表征膜结合蛋白质