STRUCTURAL AND CELL BIOLOGY IN CARDIOVASCULAR DISEASE

该计划的总体目标是了解结构 生物学以及大分子和大分子的功能 参与脂质进出细胞运输的复合物。 的 这些项目的目标是：确定 细胞受体及其配体，如LDL受体结合到 LDL计划;研究原始新生的 内质网中富含磷脂的颗粒 N端41%载脂蛋白B的二级和三级结构 规范这一过程项目;了解结构和 apo-B的构象，LDL的三维结构， 组织载脂蛋白B的LDL，并确定构象和3- 可交换载脂蛋白的空间结构 血浆HDL项目;了解白蛋白的三维结构 和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进入细胞， 与脂肪酸结合蛋白的相互作用，以及脂肪 酸对细胞内pH和钙的影响。 State-of-the-art 结构生物学技术被用来研究分离的或 重组的大分子复合物，由蛋白质或脂质组成， 蛋白质复合物和聚集体。 个体的低分辨率结构 粒子（和/或二维阵列）通过电子 显微镜，特别是冷冻EM，结合图像分析， 重建 用特定地点的标签装饰这些结构， 例如特异性表位的抗体，与游离半胱氨酸结合的金， 抗体/Fab、配体等，帮助确定具体的 大分子复合物中的结构域或序列。 以确定详细的 分子分辨率的结构，单个蛋白质或蛋白质-脂质 将配合物结晶并通过X射线衍射确定结构。 结晶学 模型特异性肽的溶液结构 蛋白质或小蛋白质区域，如细胞内脂肪酸 通过多维NMR测定结合蛋白。 高分辨率 分子排列可以叠加到低分辨率的 通过电子显微镜获得的大分子组装结构， 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， 参与脂质代谢过程的生物结构 移动到细胞内，细胞内，细胞外 这些信息将使 开发新的基于分子的策略来控制超β 脂蛋白血症、脂肪酸诱导的缺血性细胞损伤，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.