MACROMOLECULES OF CIRCULATORY AND GENETIC IMPORTANCE

具有循环和遗传重要性的大分子

• 批准号: 3268562
• 负责人: VERNE N SCHUMAKER
• 金额: $ 22.11万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3268562
• 关键词: affinity chromatography; analytical ultracentrifugation; antiidiotype antibody; apolipoproteins; blood lipoprotein; blood lipoprotein metabolism; chemical binding; chimeric proteins; chylomicrons; circular dichroism; complement pathway; conformation; electron microscopy; human tissue; immune complex; immunoglobulin G; immunoglobulin M; immunoglobulin structure; laboratory rabbit; low density lipoprotein; monoclonal antibody; protein structure function; receptor; recombinant DNA; site directed mutagenesis; stoichiometry; very low density lipoprotein; western blottings

This grant will support research in two areas: lipoprotein structure and molecular immunology. The common thread is our interest in the structure of functional macromolecular complexes formed between proteins and lipids and receptors, and between antibodies, antigen, and complement. The goals of the lipoprotein projects are to (1) determine the conformation of apolipoprotein B on the surface of the low density lipoprotein, and the location of the site on apo B recognized by the receptor; (2) to determine the number of apo B48 molecules on the chylomicrons, and to examine the confomation of apo B100 on the VLDL; (3) to characterize the structure of recombinant apolipoprotein(a) and its complex with LDL which forms the highly atherogenic Lp(a). The goals of the molecular immunology projects are (1) to determine the nature of the conformational changes through which IgM acquires the ability to bind and activate complement; (2) to develop a new technique we call 'targeted immunogenesis' to form monoclonal antibodies to selected regions on a protein. These monoclonals will be used as probes of IgM structure and function. (3) to study protein protein interactions by equilibrium dialysis, employing a special membrane not previously used for such studies. This technique will then be employed in both the lipoprotein and immunoglobulin projects. The methodology includes analytical ultracentrifugation, circular dichroism, fluorescence, and electron microscopy, as well as the production of monoclonal antibodies and recombinant DNA techniques. These projects are related to problems of health; thus (1) high concentrations of chylomicron and VLDL remnants remaining after lipolysis, and especially high concentrations of LDL, are closely associated with the development of the atherosclerotic lesion, and associated heart attack, senility and stroke; (2) the triggering of complement and cells by immune complexes and antigens is a critical feature of the immune defense against bacterial and viral challenge.

这笔赠款将支持两个领域的研究：脂蛋白结构和 分子免疫学。 共同点是我们对 蛋白质之间形成的功能性大分子复合物的结构 以及脂质和受体之间，以及抗体、抗原和 补充。 脂蛋白项目的目标是 (1) 确定 低密度表面载脂蛋白B的构象 脂蛋白，以及apo B上被识别的位点的位置 受体； (2) 测定apo B48分子数 乳糜微粒，并检查 APO B100 在 VLDL 上的构象； (3)表征重组载脂蛋白(a)的结构和 它与 LDL 形成复合物，形成高度致动脉粥样硬化的 Lp(a)。 分子免疫学项目的目标是 (1) 确定 IgM 通过构象变化的性质获得 结合和激活补体的能力； (2) 开发新技术 我们称之为“靶向免疫发生”，以形成单克隆抗体 蛋白质上的选定区域。 这些单克隆抗体将用作探针 IgM 结构和功能。 (3)研究蛋白质蛋白质相互作用 通过平衡透析，采用以前没有的特殊膜 用于此类研究。 该技术随后将应用于 脂蛋白和免疫球蛋白项目。 该方法包括分析超速离心、循环 二色性、荧光和电子显微镜，以及 单克隆抗体的生产和重组DNA技术。 这些项目与健康问题有关；因此 (1) 高 乳糜微粒和 VLDL 残留物的浓度 脂肪分解，特别是高浓度的 LDL，与 与动脉粥样硬化病变的发展有关，并且 相关的心脏病发作、衰老和中风； (2) 触发 免疫复合物和抗原对补体和细胞的作用是至关重要的 针对细菌和病毒挑战的免疫防御功能。