Nanoscale Biomembrane Characterization: Model Systems to Cells

新生物物理技术的发展对科学进步至关重要。 这项建议旨在推进现有的光谱成像技术，以阐明对细胞膜的理解，并通过开发应用于生物系统的具有前所未有的化学和空间分辨率的新的模型系统和技术来促进这种理解。拉曼近场扫描光学显微镜(NSOM)是在K99相中发展起来的，它将使振动光谱成像具有纳米级的空间分辨率。振动光谱成像提供了仅根据存在的分子在空间上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，而不需要外部标记，这可能会扰乱系统的行为。这项建议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。具体地说，将利用模型系统来探索从这一新方法中获得的生物物理见解。平面支撑的双层膜将被用作结构域形成基序的模型系统，特别是在脂类结构域中发现的化学组成和相互作用。L将开发更复杂的模型系统，如受限囊泡，这将有助于观察和理解膜的异质性及其在细胞膜过程中的作用，如信号传递和跨细胞膜的运输。最终，这项建议将利用已建立的仪器，并推进方法学，以获得对完整细胞膜的洞察。 虽然结构域通常含糊不清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，但这项研究将证明并澄清这些分子相互作用在以前无法达到的水平。

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.