Electron Crystallography of Ca++ - ATPase from Sarcoplasmic Reticulm

肌浆网Ca - ATP酶的电子晶体学

• 批准号: 9118511
• 负责人: Kenneth Taylor
• 金额: $ 24万
• 依托单位: Duke University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-15 至 1995-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9118511&HistoricalAwards=false
• 关键词: Electron; Crystallography; Ca++; ATPase; Sarcoplasmic

Ca2+ ion transport by the Ca2+-ATPase of sarcoplasmic reticulum is a fundamental process in muscle physiology. This enzyme removes Ca2+ from the cytoplasm to storage sites in the sarcoplasmic reticulum resulting in muscle relaxation. This project focusses on electron crystallographic analysis of large multilayered crystalline arrays produced from detergent solubilized enzyme in the presence of high Ca2+ concentrations. The studies proposed will 1. develop methodology for obtaining and combining electron diffraction image data from multilayered crystals of different thickness, 2. optimize the protocols for preservation of 3-D order in the crystals for electron microscopic analysis, 3. investigate crystallization protocols that favor the production of 2-D sheets free from stacking to facilitate the electron crystallographic analysis at higher resolution, and 4. obtain information on protein secondary structure of the Ca2+-ATPase molecule by searching for characteristic diffraction from alpha helices, beta sheets and lipid in X-ray diagrams of pelleted crystals. Efforts will be made to carry this structure analysis at least to the resolution of protein secondary structure (approximately 7A) to test current models of Ca2+-ATPase structure.

肌浆网Ca ~（2+）-ATP酶的Ca ~（2+）转运是肌浆网Ca ~（2+）转运的重要途径。 是肌肉生理学的基本过程 这种酶可以去除 Ca 2+从细胞质到肌浆中的储存位点 导致肌肉松弛的网状物。 该项目重点 大型多层膜的电子晶体学分析 从洗涤剂溶解的酶中产生的晶体阵列， 高浓度的Ca 2+。 建议的研究 威尔1. 开发获取和结合电子 来自不同的多层晶体的衍射图像数据 厚度，2. 优化保存3D图像的方案 电子显微镜分析的晶体中的顺序，3. 研究有利于生产的结晶方案 2-D片自由堆叠，以促进电子 更高分辨率的晶体学分析，以及4. 获得 Ca 2 +-ATPase蛋白质二级结构信息 通过搜索来自α的特征衍射， 螺旋，β折叠和脂质的X射线图中的颗粒 晶体 将努力进行这一结构分析 至少对蛋白质二级结构的解析 （约7A）以测试Ca 2 +-ATPase的当前模型 结构