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

• 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
• 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

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

总结 圆二色性（CD）和拉曼旋光性（罗阿）是提供有价值的手性光谱学。 生物分子和药物在天然条件下在水性缓冲液中的结构信息， 需要特殊的样品制备或结晶。这两种方法是互补的，因为它们探测 分子电子和分子振动跃迁的圆二色性。的组合 这两种方法特别适合于研究膜结合蛋白的结构， 仍然很难用其他生物物理表征工具表征。虽然理论上， CD /罗阿表征有可能提供膜结合的重要结构信息 蛋白质，在实践中，这两种光谱的弱灵敏度使得难以实现这种潜力。一 对高样品浓度和长采集时间的需要限制了CD的更广泛使用， 特别是作为表征膜结合蛋白工具的罗阿光谱学。该项目旨在 通过开发等离子体增强CD，克服CD和罗阿光谱的灵敏度限制 例如，使用等离子体纳米天线的PECD（PECD）和表面增强罗阿（SEROA）光谱， 具有特定电场（E）和磁场（H）特性以及限定相的工程化纳米结构 属性，以增强信号强度。为了使信号增强最大化，天线基板将 利用紫外线（UV）中的等离子体共振来开发， 与生物靶分子的分子电子共振，促进强信号强度， CD和罗阿。由于该提案侧重于开发PECD和SEROA作为表征工具， 膜结合蛋白，所提出的天线的另一个重要设计组件是组装 等离子体纳米天线表面上的脂质膜，以提供用于膜结合的结合位点 proteins.这种方法在CD和罗阿信号传导的电磁热点中富集感兴趣的蛋白质 增强是最高的，并允许光谱表征的蛋白质结构，在其 膜结合型开发的等离子体增强光谱学将使重要的新见解 膜结合蛋白的结构和手性，例如作为脂质组成的函数，和 将有助于大大提高对蛋白质-膜相互作用的理解。具体目标是 适用于： 目的1：建立膜结合蛋白质的等离子体增强紫外圆二色光谱 目的2：开发膜结合蛋白质的等离子体增强拉曼光学活性（罗阿）光谱 目标3：用于表征膜结合的原型组合电子CD /罗阿仪器 蛋白