CHARCTERIZATION OF BASEMENT MEMBRANE GLYCOPROTEINS

基底膜是复杂的细胞外基质结构 这对发展、组织和 维持组织和复杂生物体。他们在 与细胞表面紧密接触，从而影响细胞 行为的机制尚未完全理解。几 明确定义的大分子组分如IV型胶原， 层粘连蛋白、硫酸乙酰肝素蛋白聚糖和巢蛋白已经被 定位于基底膜。生物合成和组装 这种超分子基质和调节机制 在正常和患病状态下的情况需要进一步研究。的 这项建议的目的是解决这些问题。的 该提案的具体目标是： I. 分析A、B1和B2的组装规律 层粘连蛋白链; 二. 研究层粘连蛋白-巢蛋白复合物的组装。 为了实现这些目标，组装过程将 在完整细胞和无细胞系统中进行了研究。 抗体 将获得对每种分子特异的用于分析。 含有各层粘连蛋白链的cDNA序列的载体 将被构建并用于干扰细胞内 层粘连蛋白的组装。 稳定的细胞系，其产生修饰的 将获得并研究矩阵。 的相互作用 层粘连蛋白和巢蛋白可能在调节 基底膜功能和特异性。 埃森将是 在杆状病毒表达系统中产生的， 检查层粘连蛋白的组成。 此外该 层粘连蛋白-巢蛋白复合物的细胞内组装将是 通过用载体构建体转染来调节它来研究 含有巢蛋白cDNA序列。 这项工作的最终目标是了解地下室 膜组装，是什么调节组装，以及如何组装， 基底膜功能在健康和疾病中特异性 与这些过程有关。

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.