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CHARACTERIZATION OF MEMBRANE LIPID DOMAINS

膜脂域的表征

基本信息

批准号：
3291250
负责人：
RUTH WELTI
金额：
$ 9.55万
依托单位：
KANSAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-03-01 至 1991-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3291250
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

项目摘要

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，脂质都有能力存在于不同于其他横向域的横向域中作文中的分组。然而，这一安排生物膜内的磷脂分子物种不是很好理解。在拟议的工作中，化学成分合成脂质混合物中的脂相和脂结构域，分离的大肠杆菌脂类和完整的细胞质将对大肠杆菌的细胞膜进行研究。荧光将使用对位芳酰基磷脂的极化来评估脂肪相存在，而最近的化学交联邻近的脂质将被用来确定脂质的排列膜液相脂。磷脂酰乙醇胺物种将通过胺特定的试剂进行交联，二甲基硫酰亚胺及其高压分析产物液体和气-液色谱。脂类物种，映射到一个完整膜中的相和结构域，将定位于使用非特异性脂质转移的内或外单分子层牛肝中的蛋白质及其化学标记三硝基苯磺酸。这些研究将考察脂肪酸代谢发生突变的大肠杆菌的细胞膜。在各种脂肪酸补充剂的作用下生长，这些细胞由于它们的脂类而产生功能缺陷。作文。这里描述的化学和物理研究将集成到一个更清晰定义的膜模型中结构，从长远来看，这将提供一个联系的基础膜脂拓扑结构对膜功能的影响。