STUDIES OF LIPID-PROTEIN AND PROTEIN-PROTEIN INTERACTIONS IN HIV

HIV 中脂质-蛋白质和蛋白质-蛋白质相互作用的研究

• 批准号: 3939287
• 负责人: J N WEINSTEIN
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3939287
• 关键词: T lymphocyte; atherosclerosis; blood lipoprotein metabolism; chromatography; drug interactions; electron microscopy; fluorescent dye /probe; high density lipoproteins; human immunodeficiency virus 1; human tissue; image processing; immunological substance; liposomes; membrane lipids; membrane reconstitution /synthesis; membrane structure; neutron diffraction; stoichiometry; surface antigens

We have investigated the interaction of lipoproteins with liposomes to form recombinant particles. A number of lipoprotein fractions (VLDL, IDL, LDL, and HDL) all disrupt liposome structure by an essentially irreversible and quasistoichiometric process. In the case of HDL, the major apoprotein, A-I, recombines with dimyristoyl phosphatidyl choline vesicles 40:1 lipid-protein to form discs approximately 100 angstrom unit in diameter and 32 angstrom unit in thickness, with protein on the rim. These structural results were obtained by a combination of neutron scattering, electron microscopy, column chromatography, and fluorescence techniques. With dipalmitoyl phosphatidylcholine, A-I also forms what we term "vesicular recombinant" particles in a process which may relate to physiological mechanisms by which proteins are assembled into membranes and lipoproteins. To study this process we have developed a technique called "phase transition release" (PTR) which is also being applied to study incorporation of tubulin into membranes. Lipoproteins were labelled with the fluorescent lipid 3,3 dioctadecylindo-carbocyanine for studies of interaction will cell surface lipoprotein receptors. The lipoproteins are also being labelled with NBD lipids for two-color fluorescence identification of cells in atheroscleroic plaques. Statistical and more general mechanical algorithms (HAL, HALP, HALCO) were devised for evaluating amphipathic helical structures and more general structure-function relationships in proteins and peptides. This is being used to define issues of structure and immunogenicity with respect to HLA antigens and to the envelope polyprotein of HIV. Lipid membrane systems and human cell isolates are being used experimentally to investigate the interaction between characterized synthetic antigenic peptides and T-cells in the recognition process. The results may have application to the design of vaccines.

我们研究了脂蛋白与 脂质体以形成重组颗粒。 一些脂蛋白 组分（VLDL、IDL、LDL和HDL）均破坏脂质体 通过基本上不可逆和准化学计量的 过程 在HDL的情况下，主要的载脂蛋白A-I， 与二肉豆蔻酰磷脂酰胆碱囊泡40：1重组 脂质-蛋白质形成约100埃单位的圆盘， 直径和32埃单位的厚度，与蛋白质上 边缘。 这些结构结果是通过以下组合获得的： 中子散射，电子显微镜，柱层析， 和荧光技术。 与二棕榈酰磷脂酰胆碱，A-I也形成了我们 术语“囊泡重组体”颗粒， 与蛋白质在人体内 组装成膜和脂蛋白。 研究这一过程 我们开发了一种叫做“相变释放”的技术 (PTR)这也被应用于研究微管蛋白的结合 变成膜。 用荧光脂质3，3标记脂蛋白 双十八烷基吲哚碳菁与细胞相互作用的研究 表面脂蛋白受体 脂蛋白也被 用NBD脂质标记，用于双色荧光鉴定 动脉粥样硬化斑块中的细胞。 统计和更一般的机械算法（HAL，HALP， HALCO）设计用于评价两亲性螺旋 结构和更一般的结构-功能关系， 蛋白质和肽。 这被用来定义 结构和免疫原性， HIV的包膜多聚蛋白。 脂质膜系统和 人类细胞分离物正在实验性地用于研究 特征性合成抗原之间的相互作用 肽和T细胞在识别过程中。 结果可能 可以应用于疫苗的设计。