STRUCTURAL CELL BIOLOGY IN CARDIOVASCULAR DISEASE

心血管疾病中的结构细胞生物学

• 批准号: 6627520
• 负责人: DAVID ATKINSON
• 金额: $ 224.53万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-30 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6627520
• 关键词: blood lipoprotein; cardiovascular disorder; molecular pathology; protein structure; structural biology

The objective of this program is to understand the Structural Biology of the macromolecular complexes involved in the transport of lipids into and out of cells. The aims of the project are: to define the 3-dimensional structures of cellular receptors and their ligands bound to LDL (Project 1): to study the formation of primordial nascent triglyceride-rich particles in the endoplasmic reticulum and understand how the secondary and tertiary structures of the N-terminal 41% apo-B regulates this process (Project 2); to understand the structure and conformation of apo-B, the 3- dimensional structure of LDL, the organization of apo-B on LDL, and to define the formation and 3-dimensional structure of exchangeable apolipoproteins on nascent ans plasma HDL (Project 3); to understand the binding of fatty acids to albumin, the transfer of fatty acids from albumin and movement across membranes into cells, structural and dynamic features of intracellular fatty acid binding proteins, and the effect of fatty acids on intracellular Ph (Project 4). State-of-the-art techniques of structural biology are used to study isolated or reconstituted macromolecular complexes. Low resolution structures of individual particles (and/or 2 dimensional arrays) often decorated with site specific labels are obtained by electron microscopy, in particular cryo-EM, combined with image analysis and reconstruction. Detailed structures of individual proteins or protein-lipid complexes are determined by x-ray crystallography. The solution structure of peptides that model regions of proteins or small proteins such as the intracellular fatty acid binding proteins are determined by multi-dimensional NMR. High resolution molecular arrangements can be superimposed onto the low resolution structure of macromolecular assemblies obtained by electron microscopy to generate a "higher resolution" macromolecular structure. These studies can provide the biological structures involved in the processes by which lipids are moved into, within, and out of cells. Such information will allow the development of new molecular based strategies to control hyper beta lipoproteinemia, fatty acid-induced cellular damage in ischemia, and arteriosclerosis.

本课程的目的是了解参与脂质进出细胞运输的大分子复合物的结构生物学。该项目的目标是：定义细胞受体及其与LDL结合的配体的三维结构（项目1）；研究内质网中原始新生富甘油三酯颗粒的形成，并了解n端41%载脂蛋白b的二级和三级结构如何调节这一过程（项目2）；了解载脂蛋白b的结构和构象，LDL的三维结构，LDL上载脂蛋白b的组织，确定新生体和血浆HDL上可交换载脂蛋白的形成和三维结构（项目3）；了解脂肪酸与白蛋白的结合，脂肪酸从白蛋白转移并跨膜进入细胞，细胞内脂肪酸结合蛋白的结构和动态特征，以及脂肪酸对细胞内Ph的影响（项目4）。最先进的结构生物学技术被用于研究分离的或重组的大分子复合物。通过电子显微镜，特别是低温电镜，结合图像分析和重建，可以获得具有特定位点标记的单个粒子（和/或二维阵列）的低分辨率结构。单个蛋白质或蛋白质-脂质复合物的详细结构由x射线晶体学确定。模拟蛋白质区域或小蛋白质（如细胞内脂肪酸结合蛋白）的肽的溶液结构是由多维核磁共振确定的。高分辨率分子排列可以叠加到由电子显微镜获得的低分辨率大分子组合结构上，从而产生“更高分辨率”的大分子结构。这些研究可以提供与脂质进入、进入和离开细胞的过程有关的生物结构。这些信息将允许开发新的基于分子的策略来控制高β脂蛋白血症、脂肪酸诱导的缺血细胞损伤和动脉硬化。