CHARACTERIZATION OF MEMBRANE LIPID DOMAINS

膜脂域的表征

• 批准号: 3291249
• 负责人: RUTH WELTI
• 金额: $ 9.37万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-03-01 至 1990-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3291249
• 关键词: Escherichia coli; chemical structure function; crosslink; fluorescence polarization; gas chromatography; high performance liquid chromatography; intracellular membranes; membrane lipids; membrane model; membrane reconstitution /synthesis; membrane structure; microorganism culture; phosphatidylethanolamines; phospholipids; reagent /indicator

In both model and biological membranes, lipids have the capability of existing in lateral domains which differ from other lateral groupings in composition. However, the arrangement of phospholipid molecular species within biological membranes is not well understood. In the proposed work, the chemical composition of lipid phases and lipid domains in synthetic lipid mixtures, isolated Escherichia coli lipids, and the intact cytoplasmic membrane of E. coli will be investigated. Fluorescence polarization of parinaroyl phospholipids will be used to assess the lipid phases present, while chemical crosslinking of nearest neighbor lipids will be used to determine the lipid arrangement of the membrane fluid phase lipids. Phosphatidylethanolamine species will be crosslinked by the amine-specific reagent, dimethylsuberimidate, and the products analyzed by high pressure liquid and gas-liquid chromatography. Lipid species, mapped to a phase and domain in intact membranes, will be localized to the inner or outer monolayer using the nonspecific lipid transfer protein from bovine liver and by chemical labelling with trinitrobenzenesulfonic acid. These studies will examine the membranes of E. coli with mutations in fatty acid metabolism. Grown with various fatty acid supplements, the membranes of these cells develop functional defects due to their lipid compositions. The chemical and physical studies described here will be integrated into a more clearly defined model of membrane structure, which, in the long term, will provide a basis to relate membrane lipid topology to membrane function.

在模型膜和生物膜中，脂质都具有以下能力 存在于与其他横向​​域不同的横向域中的 组成上的分组。 然而，安排 生物膜内的磷脂分子种类不是 很好理解。 在拟议的工作中，化学成分 合成脂质混合物中的脂质相和脂质域， 分离的大肠杆菌脂质和完整的细胞质 将研究大肠杆菌的膜。 荧光 仲芳酰磷脂的极化将用于评估 存在脂质相，而最近的化学交联 相邻脂质将用于确定脂质排列 膜流体相脂质。 磷脂酰乙醇胺 物种将被胺特异性试剂交联， 辛二酰亚胺酸二甲酯及高压分析产物 液相色谱和气液色谱。 脂质种类，映射到 完整膜中的相和域，将定位于 使用非特异性脂质转移的内部或外部单层 来自牛肝的蛋白质并通过化学标记 三硝基苯磺酸。 这些研究将检验 脂肪酸代谢发生突变的大肠杆菌膜。 在添加各种脂肪酸补充剂的情况下生长的细胞膜 这些细胞由于其脂质而出现功能缺陷 组成。 这里描述的化学和物理研究 将被整合到一个更清晰定义的膜模型中 的结构，从长远来看，这将为联系提供基础 膜脂质拓扑结构与膜功能的关系。