SPECTROSCOPY OF BIOMEMBRANE MONOLAYERS

单层生物膜的光谱学

• 批准号: 2903173
• 负责人: RICHARD A. DLUHY
• 金额: $ 26.79万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2903173
• 关键词: Raman spectrometry; bioimaging /biomedical imaging; biophysics; conformation; fiber optics; fluorescence microscopy; hydropathy; infrared spectrometry; intermolecular interaction; light scattering; lipid structure; membrane model; membrane proteins; membrane structure; molecular film; phase change; phospholipids; protein structure; pulmonary surfactants; surface property

It is the overall aim of the current proposal to employ a combined spectroscopy/imaging approach using IR and Raman spectroscopy as well as fluorescence microscopy for the study of biophysical monolayer models of pulmonary surfactant. A combined approach is needed, since no one method is able to accomplish both structural characterization and morphological imaging for a detailed understanding of conformational and spatial properties in the monolayer. The combination of vibrational spectroscopy and fluorescence imaging will correlate molecular properties with mesoscopic (i.e. mum size) domain morphology in surfactant monolayers. Our recent results suggest that the interaction of surfactant proteins with biomembrane interfaces may occur selectively at locations with distinct morphological properties, such as at domain boundaries or areas of a certain lipid phase. Therefore, investigation of the molecular structural properties of the system, as reported by IR or Raman, in conjunction with its morphological structure, as visualized by fluorescence, is needed to more fully understand the interaction between the system components. The combination of techniques described in this proposal will mark an important achievement for the investigation of the molecular interaction between proteins and lipids in surfactant models in that we will be able to associate specific molecular spectroscopic properties with lateral organization in surface monolayers. During the next grant period, we will address the following specific aims: 1) to characterize the interfacial and hydrophobic interactions of the surfactant proteins SP-B and SP-C on monolayer structure by IR and to determine protein structure in the monolayer by Raman spectroscopy, 2) to characterize the influence of SP-B and SP-C on monolayer morphology by fluorescence microscopy and to determine how these proteins produce three-dimensional, surface-excluded material as seen using scattered light microscopy, 3) to use IR spectroscopy to measure microphase separation and domain formation in binary mixtures, and 4) to use spatially-resolved Raman microscopy to study lateral organization and molecular structure in surfactant monolayer films.

目前建议的总体目标是采用红外和拉曼光谱以及荧光显微镜研究肺表面活性物质的生物物理单层模型的组合光谱/成像方法。 需要一种组合的方法，因为没有一种方法能够同时完成结构表征和形态成像，以详细了解单层中的构象和空间性质。 振动光谱和荧光成像的组合将与表面活性剂单分子膜中的介观（即妈妈大小）域形态的分子特性。 我们最近的研究结果表明，表面活性剂蛋白与生物膜界面的相互作用可能会选择性地发生在具有不同形态学特性的位置，例如在域边界或某些脂质相的区域。因此，需要研究该系统的分子结构特性，如IR或拉曼所报道的，结合其形态结构，如荧光所显示的，以更充分地理解系统组分之间的相互作用。 在这个建议中描述的技术的组合将标志着一个重要的成就，在表面活性剂模型中的蛋白质和脂质之间的分子相互作用的调查，我们将能够关联特定的分子光谱特性与表面单分子层的横向组织。 在下一个资助期内，我们会致力达致以下具体目标：1）通过IR表征表面活性剂蛋白SP-B和SP-C在单层结构上的界面和疏水相互作用，并通过拉曼光谱确定单层中的蛋白质结构，2）通过荧光显微镜来表征SP-B和SP-C对单层形态的影响，并确定这些蛋白质如何产生三个-三维的，表面排除的材料，如使用散射光显微镜，3）使用红外光谱测量微相分离和域形成的二元混合物，和4）使用空间分辨拉曼显微镜研究横向组织和分子结构的表面活性剂单层膜。