CHARACTERIZATION OF BASEMENT MEMBRANE GLYCOPROTEINS

基底膜糖蛋白的表征

• 批准号: 3273248
• 负责人: ALBERT E CHUNG
• 金额: $ 18.09万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3273248
• 关键词: basement membrane; cell free system; complementary DNA; entactin; extracellular matrix; gene expression; genetic translation; glycoprotein biosynthesis; glycoproteins; immunochemistry; insect virus; laboratory mouse; laminin; membrane proteins; membrane reconstitution /synthesis; membrane structure; messenger RNA; molecular cloning; monoclonal antibody; posttranslational modifications; protein biosynthesis; protein reconstitution; protein sequence; protein structure; protein structure function; tissue /cell culture; transfection; transposon /insertion element

Basement membranes are complex extracellular matrix structures that are essential for the development, organization, and maintenance of tissues and complex organisms. They are in intimate contact with cell surfaces and thereby influence cell behavior by mechanisms that are not fully understood. Several well-defined macromolecular components such as type IV collagen, laminin, heparan sulfate proteoglycan and entactin have been localized to the basement membrane. The biosynthesis and assembly of this supra-molecular matrix and the mechanisms which regulate them in normal and diseased states require further study. The objective of this proposal is to address those problems. The specific aims of the proposal are: I. To analyze the regulation of assembly of the A, B1 and B2 chains of laminin; II. To study the assembly of the laminin-entactin complex. To accomplish these objectives the assembly process will be studied in both intact cells and cell-free systems. Antibodies specific for each molecule will be obtained for the analyses. Vectors containing cDNA sequences for each of the laminin chains will be constructed and used to perturb the intracellular assembly of laminin. Stable cell lines which produce modified matrices will be obtained and studied. The interaction of laminin and entactin may play an important role in regulating basement membrane function and specificity. Entactin will be produced in the baculovirus expression system and its re- constitution with laminin will be examined. In addition, the intracellular assembly of the laminin-entactin complex will be studied by modulating it by transfection with vector constructs containing entactin cDNA sequences. The ultimate goal of this work is to understand how basement membranes are assembled, what regulates the assembly, and how the specificity of basement membrane function in health and disease is related to these processes.

地下膜是复杂的细胞外基质结构 这对于发展，组织和 维持组织和复杂生物。他们在 与细胞表面的亲密接触，从而影响细胞 通过未完全理解的机制行为。一些 定义明确的大分子成分，例如IV型胶原蛋白， 层粘连蛋白，硫酸乙酰肝素蛋白聚糖和Entactin是 位于地下室膜。生物合成和组装 该上分子基质和调节的机制 在正常和患病状态下，他们需要进一步研究。这 该提案的目的是解决这些问题。这 该提案的具体目的是： I.分析A，B1和B2组装的调节 层粘连蛋白的链； ii。 研究层粘连蛋白 - 肠菌素复合物的组装。 为了实现这些目标，组装过程将是 在完整的细胞和无细胞系统中进行了研究。 抗体 对于分析，将获得每个分子的特异性。 每个层粘连蛋白链中含有cDNA序列的载体 将构建并用于扰动细胞内 层粘连蛋白的组装。 产生修饰的稳定细胞系 将获得和研究矩阵。 相互作用 层粘连蛋白和Entactin可能在调节中发挥重要作用 地下室膜功能和特异性。 Entactin将是 在杆状病毒表达系统中产生及其重新 将检查使用层粘连蛋白的宪法。 另外， 层粘连蛋白 - 肠菌素复合物的细胞内组件将是 通过通过向量构建体转染来调节它来研究它 包含肠蛋白cDNA序列。 这项工作的最终目标是了解地下室 膜是组装的，是什么调节组件，以及如何 地下室膜功能在健康和疾病中的特异性 与这些过程有关。