Nanoscale Biomembrane Characterization: Model Systems to Cells

纳米级生物膜表征：细胞模型系统

• 批准号: 7810125
• 负责人: Zachary Schultz
• 金额: $ 24.89万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7810125
• 关键词: Agitation; Area; Behavior; Binding; Biological; Biological Assay; Biological Models; Caliber; Carbon; Carrier Proteins; Cell membrane; Cells; Cellular Membrane; Characteristics; Charge; Chemicals; Cholesterol; Color; Complex; Computers and Advanced Instrumentation; Controlled Environment; Development; Divalent Cations; Electrostatics; Future; Generations; Goals; Gold; Heterogeneity; Image; Imaging Techniques; Infection; Investigation; Label; Lateral; Light; Lipid Bilayers; Lipids; Location; Magnesium; Manuscripts; Maps; Measurement; Measures; Membrane; Methodology; Methods; Microscope; Optics; Organelles; Performance; Phase; Photoreceptors; Preparation; Process; Property; Proteins; Publications; Radial; Ramp; Reproducibility; Research Activity; Resolution; Rhodopsin; Rod Outer Segments; Role; Sampling; Scanning; Scanning Probe Microscopes; Science; Sedimentation process; Signal Transduction; System; Techniques; Temperature; Tissues; Ultrasonics; Velocimetries; Vesicle; Work; base; biological systems; cell behavior; design; infrared spectroscopy; insight; light scattering; molecular domain; nanoparticle; nanoscale; pathogen; retinal rods; spectroscopic imaging; submicron

The development of new biophysical techniques is of paramount importance to the advancement of science. This proposal aims to advance existing spectroscopic imaging techniques to elucidate the understanding of cellular membranes and to further this understanding by developing both new model systems and techniques with unprecedented chemical and spatial resolution for application to biological systems. A Raman near fleld scanning optical microscope (NSOM) was developed during the K99 phase that will enable vibrational spectroscopic imaging with nanoscale spatial resolution. Vibrational spectroscopic imaging offers the opportunity to spatially resolve biological systems solely on the basis ofthe molecules present. This enables the study of the dynamical and organizational characteristics of cells and tissue without the need for external labels, which can disrupt the behavior of the system. This proposal aims to further develop and apply these new techniques to elucidate further the characteristics of biomolecules located within cellular membranes. Specifically, model systems will be utilized to explore the biophysical insights gained from this new methodology. Planar supported bilayers will be used as a model system for domain formation motifs, particularly the chemical composition and interactions that are found in lipid domains. l\/Iore sophisticated model systems, such as confined vesicles, will be developed that will facilitate the ultimate goal of observing and understanding membrane heterogeneity and its role in cellular membrane processes such as signaling, and transport across the cell membrane. Ultimately, this proposal will utilize the established instrumentation and advance the methodology to gain insight into intact cell membranes. While domains are commonly ambiguously implicated in cell membrane processes related to pathogen infection, transport, and protein receptor activity, this research will demonstrate and clarify these molecular interactions at levels previously unattainable.

新生物物理技术的发展对于科学的进步至关重要。 该提案旨在推进现有的光谱成像技术，以阐明对细胞膜的理解，并通过开发具有前所未有的化学和空间分辨率的新模型系统和技术应用于生物系统来进一步加深这种理解。 K99 阶段开发了拉曼近场扫描光学显微镜 (NSOM)，它将实现具有纳米级空间分辨率的振动光谱成像。振动光谱成像提供了仅根据存在的分子来空间解析生物系统的机会。这使得研究细胞和组织的动态和组织特征成为可能，而无需外部标签，因为外部标签可能会破坏系统的行为。该提案旨在进一步开发和应用这些新技术，以进一步阐明位于细胞膜内的生物分子的特征。具体来说，模型系统将用于探索从这种新方法中获得的生物物理见解。平面支持的双层将用作结构域形成基序的模型系统，特别是在脂质结构域中发现的化学成分和相互作用。将开发复杂的模型系统，例如受限囊泡，这将有助于实现观察和理解膜异质性及其在细胞膜过程（例如信号传导和跨细胞膜运输）中的作用的最终目标。最终，该提案将利用现有的仪器并改进方法来深入了解完整的细胞膜。 虽然结构域通常模糊地涉及与病原体感染、运输和蛋白质受体活性相关的细胞膜过程，但这项研究将以以前无法达到的水平证明和澄清这些分子相互作用。