PHYSIOCOCHEMICAL FEATURES OF LYMPHOCYTE PLASMA MEMBRANES

这个项目的长期目标是探索 B淋巴细胞血浆的物理化学组织 并了解该组织中的变化如何 反映和/或调节细胞激活的主要事件。这个 该项目的第一部分涉及组织和分配 膜结构域上的脂类。大小、脂类成分和 将使用荧光检测脂类结构域的流动性 光漂白恢复与扫描荧光相关 光谱学。大分子单层囊泡的制备及性能研究 B细胞膜脂质将有助于评估细胞脂质 组织依赖于细胞表面蛋白的存在。 通过特定的脂质浓缩引起的膜脂改变 通过糖皮质激素治疗来消耗胆固醇，以及 通过温度摄动将被检查。第二 工作领域涉及以下方面的分布和相互作用 膜蛋白。扫描荧光相关 光谱学实验将探索蛋白质是否 定位于一种或多种类型的富含蛋白质的区域 特定的脂类是否优先与 特定的蛋白质。特定的关联发生在 已知的B细胞膜蛋白和其他已知或尚未知晓的 将使用荧光能量对未知物种进行评估 转移和新的免疫化学靶向光近似性 标签方法。特异膜的聚集状态 蛋白质将使用时间分辨磷光进行评估 各向异性。最后，B淋巴细胞的即刻变化 三次活化产生的膜结构 通过抗IgM治疗，通过胸腺非依赖性的机制 类抗原配体与人工修饰的细胞相互作用 与特定的受体结合，并通过脂多糖 检查过了。

The long-range goal of this project is to explore the physicochemical organization of the B lymphocyte plasma membrane and to learn how changes in this organization may reflect and/or mediate primary events in cellular activation. The first part of this project concern the organization and distribution of lipids on membrane domains. Sizes, lipid composition, and fluidities of lipid domains will be examined using fluorescence photobleaching recovery and scanning fluorescence correlation spectroscopy. Studies of large unilamellar vesicles prepared from B cell membrane lipid will help assess whether cellular lipid organization is dependent on the presence of cell surface protein. Membrane lipid alteration via specific lipid enrichment, via cholesterol depletion through glucocorticoid treatment, and through temperature perturbation will be examined. The second area of work concerns the distribution and interaction of membrane proteins. Scanning fluorescence correlation spectroscopy experiments will explore whether proteins are localized into one or more types of protein-rich regions and whether specific lipid classes preferentially associate with particular proteins. Specific associations occurring between known B cell membrane proteins and other known or as-yet unidentified species will be assessed using fluorescence energy transfer and new immunochemically-targeted photoproximity labeling methods. The aggregation state of specific membrane proteins will be evaluated using time-resolved phosphorescence anisotropy. Finally, immediate changes in B lymphocyte membrane structure resulting from activation by three mechanisms, by anti-IgM treatment, by thymus-independent antigen-like ligands interacting with cells artificially decorated with specific receptors, and by lipopolysaccharide will be examined.