STRUCTURAL AND CELL BIOLOGY IN CARDIOVASCULAR DISEASE

本课程的总体目标是了解结构 生物学，因此，大分子和大分子的功能 参与脂类进出细胞运输的复合体。这个 这些项目的目标是：定义 细胞受体及其配体，如结合到 低密度脂蛋白项目；研究原始新生的形成 了解内质网中富含甘油三酯的颗粒 41%apo-B的N-末端的二级和三级结构 规范流程项目；了解流程项目的结构和 载脂蛋白B的构象，低密度脂蛋白的三维结构， 载脂蛋白B在低密度脂蛋白上的组织，并确定其构象和3- 新生和幼体可交换载脂蛋白的空间结构 血浆高密度脂蛋白计划：了解白蛋白的三维结构 及其特定的脂肪酸结合域，脂肪酸的转移 从白蛋白和跨膜运动到细胞，它们的结果 与脂肪酸结合蛋白的相互作用以及脂肪的影响 酸对细胞内pH和钙计划的影响。最先进的 结构生物学技术被用来研究孤立的或 重组的大分子复合体，由蛋白质或脂质组成- 蛋白质复合体和聚集体。个体的低分辨率结构 粒子(和/或二维阵列)是通过电子获得的 显微镜，特别是低温电子显微镜，结合图像分析和 重建。用特定地点的标签装饰这些建筑， 例如针对特定表位的抗体，与游离半胱氨酸结合的金， 抗体/Fbs、配体等有助于确定特定的 大分子复合体中的结构域或序列。确定详细信息的步骤 分子分辨的结构，单个蛋白质或蛋白质-脂 配合物被结晶，其结构由x射线确定 结晶学。模拟特异性的多肽的溶液结构 蛋白质或小蛋白质的区域，如细胞内脂肪酸 结合蛋白由多维核磁共振确定。高分辨率 分子排列可以叠加在低分辨率上 通过电子显微镜获得的大分子组件的结构 产生“更高分辨率”的大分子结构。这些研究 将提供基本的分子信息 脂类代谢过程中涉及的生物结构 移入、移入、移出牢房。这些信息将允许 基于分子的控制超β反应的新策略的发展 脂蛋白血症，脂肪酸在缺血时引起的细胞损伤，以及 动脉硬化。

The overall objective of this program is to understand the Structural Biology and hence, function of the macromolecules and macromolecular complexes involved in the transport of lipids into and out of cells. The aims of the projects are: to define the 3-dimensional structures of cellular receptors and their ligands, such as the LDL receptor bound to the LDL Project; to study the formation of primordial nascent triglyceride-rich particles in the endoplasmic reticulum and understand how the secondary and tertiary structure of the N-terminal 41% apo-B regulates this process Project; 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 conformation and 3- dimensional structure of exchangeable apolipoproteins on nascent and plasma HDL Project; to understand the 3-dimensional structure of albumin and its specific fatty acid-binding domains, the transfer of fatty acids from albumin and movement across membranes into cells, their consequent interaction with fatty acid binding proteins, and the effect of fatty acids on intracellular pH and calcium Project. State-of-the-art techniques of structural biology are used to study isolated or reconstituted macromolecular complexes, consisting of proteins or lipid- protein complexes and aggregates. Low resolution structures of individual particles (and/or 2-dimensional arrays) are obtained by electron microscopy, in particular cryo-EM, combined with image analysis and reconstruction. Decorating these structures with site-specific labels, such as antibodies to specific epitopes, gold bound to free cysteine, antibody/Fabs, ligand, etc., aids in defining the position in specific domains or sequences in the macromolecular complex. To determine detailed structures at molecular resolution, individual proteins or protein-lipid complexes are crystallized and the structure determined by x-ray crystallography. The solution structure of peptides that model specific 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 will provide fundamental molecular information essential to understanding 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.