LIPOPROTEIN STRUCTURE AND APOPROTEIN CONFORMATION

脂蛋白结构和脱辅基蛋白构象

• 批准号: 6302137
• 负责人: DAVID ATKINSON
• 金额: $ 34.7万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302137
• 关键词: X ray crystallography; apolipoproteins; blood chemistry; blood lipoprotein; calorimetry; chemical stability; circular dichroism; conformation; electron microscopy; endopeptidases; gel filtration chromatography; high performance liquid chromatography; human tissue; kallikreins; lipid structure; molecular dynamics; monoclonal antibody; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; protein structure; structural biology; synthetic peptide; thermodynamics; thrombin

The major objectives of this project are to provide detailed structural, conformational and dynamic information on the molecular organization and interactions of lipids and apoproteins in the plasma lipoproteins (primarily HDL and LDL). Studies focus on high resolution structural investigations of specific apoproteins and lipoproteins using state-of- the-art methodologies of structural biology. The primary areas explored concern: a. the three dimensional structure of intact lipoproteins (HDL, LDL) with emphasis on the molecular conformation of the apoproteins of the lipoprotein surface, and b. the detailed molecular conformation and structure of the apolipoproteins (AI and B). Studies of apolipoproteins AI will employ a combination of methodologies for the identification of important structural and functional elements in the sequence. Synthetic peptides will be used to model these regions. The detailed molecular conformation and thermodynamic stability will be studied by modelling, X- ray crystallography, 2D NMR, CD, and calorimetry. In addition the detailed molecular structure of the native apoproteins will be studied by X-ray crystallography. Studies of apoprotein B will focus on the domain folding of the protein and the conformation and function of the subdomains. Protease cleavage and molecular biology approaches will be used to obtain segments of the apoB sequence representing these structural domains. Structural studies of native LDL, by cryo electron microscopy will focus on the organization of apoB and the localization of structural and functional domains on the LDL particle using a combination of immuno- nanogold labelling. A detailed molecular understanding of apoprotein conformation and structure is vital to further progress in understanding lipoprotein structure and function. The information derived in this project will result in an improved understanding of the molecular organization and interactions in HDL and LDL. More importantly, they will provide basic background information essential to understanding physiological and metabolic processes such as lipoprotein cell surface interactions, lipoprotein interconversion and catabolism at a truly molecular level.

该项目的主要目标是提供详细的结构、 有关分子组织的构象和动态信息 血浆脂蛋白中脂质和脱辅基蛋白的相互作用 （主要是高密度脂蛋白和低密度脂蛋白）。 研究重点是高分辨率结构 使用状态研究特定的脱辅基蛋白和脂蛋白 结构生物学的最先进方法。 探索的主要领域 关注点：a.完整脂蛋白（HDL、 LDL），重点是脱辅基蛋白的分子构象 脂蛋白表面，和b．详细的分子构象和 载脂蛋白（AI 和 B）的结构。 载脂蛋白的研究 人工智能将采用多种方法的组合来识别 序列中重要的结构和功能元素。 合成的 肽将用于对这些区域进行建模。 详细分子 构象和热力学稳定性将通过建模、X- 射线晶体学、2D NMR、CD 和量热法。 此外 天然脱辅基蛋白的详细分子结构将由以下人员研究 X射线晶体学。 脱辅基蛋白B的研究将集中在以下领域 蛋白质的折叠及其构象和功能 子域。 蛋白酶裂解和分子生物学方法将 用于获得代表这些结构的 apoB 序列片段 域。 通过冷冻电子显微镜研究天然 LDL 的结构 将重点关注apoB的组织和结构的本地化 和 LDL 颗粒上的功能域，使用免疫组合 纳米金标签。 对脱辅基蛋白的详细分子了解 构象和结构对于进一步理解至关重要 脂蛋白的结构和功能。 在此得出的信息 该项目将加深对分子的理解 HDL 和 LDL 的组织和相互作用。 更重要的是，他们将 提供理解所必需的基本背景信息 生理和代谢过程，例如脂蛋白细胞表面 相互作用、脂蛋白相互转化和分解代谢 分子水平。