CALCIUM DEPENDENT MEMBRANE BINDING PROTEINS

钙依赖性膜结合蛋白

• 批准号: 2902600
• 负责人: CARL E CREUTZ
• 金额: $ 25.85万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2902600
• 关键词: Caenorhabditis elegans; Paramecium; X ray crystallography; annexins; biophysics; calcium; calcium binding protein; cell membrane; chemical binding; crystallization; electron crystallography; gene expression; green fluorescent proteins; membrane activity; membrane fusion; membrane lipids; membrane model; membrane structure; phospholipids; protein protein interaction; protein structure function; site directed mutagenesis; structural biology; synaptotagmin; yeast two hybrid system

DESCRIPTION (Adapted from abstract): Calcium-dependent, phospholipid-binding proteins associate with biological membranes in response to cell activation and calcium entry into the cytoplasm. The goal of this project is to determine the structures and biological activities of two ubiquitous classes of these proteins, the annexins, and C2-domain containing proteins. X-ray crystallography will be used to determine the structures of annexin II, the cytoplasmic domain of synaptotagmin, and the copines. Electron microscopy of 2-dimensional crystals of these proteins on lipid monolayers will be used to determine the physical nature of the interaction of these proteins with membranes. The role of individual protein domains in promoting lipid interaction and specificity will be determined by mutagenesis of the proteins and studies on model membrane systems. Proteins that interact with and are regulated by the annexins and copines will be characterized. The localization and movements of these membrane-binding proteins in living cells and tissues will be determined. The functions of the annexins and copines will be studied by genetic silencing methods in two model organisms, the nematode C. elegans, and the ciliate Paramecium tetaurelia. These peripheral proteins are hypothesized to mediate or regulate events occurring on membrane surfaces in stimulated cells. Therefore, these studies should provide insights into basic mechanisms underlying the regulation of membrane trafficking and signal transduction across membranes occurring in normal cells and in pathological conditions including cancer, neurogenic hypertension, diabetes and other endocrine disorders.

描述（改编自摘要）：钙依赖性、磷脂结合 蛋白质响应于细胞活化而与生物膜结合， 钙进入细胞质。该项目的目标是确定 结构和生物活性的两个普遍存在的类，这些 蛋白质、膜联蛋白和含C2结构域的蛋白质。x射线 晶体学将用于确定膜联蛋白II的结构， 突触结合蛋白的胞质结构域和copines。电镜观察 2-这些蛋白质在脂质单层上的三维晶体将用于 确定这些蛋白质与 膜。单个蛋白质结构域在促进脂质代谢中的作用 相互作用和特异性将通过蛋白质的诱变来确定 和模型膜系统的研究。蛋白质相互作用， 将表征由膜联蛋白和辅蛋白调节的蛋白质。国产化 以及这些膜结合蛋白在活细胞和组织中的运动 将被确定。膜联蛋白和辅蛋白的功能将被研究 在两种模式生物中，线虫C.优雅， 和纤毛草履虫。这些外周蛋白质是 假设介导或调节发生在膜表面的事件， 刺激细胞因此，这些研究应该提供基本的见解， 膜运输和信号调节的潜在机制 在正常细胞和病理细胞中发生的跨膜转导 包括癌症、神经源性高血压、糖尿病和其他 内分泌失调