MECHANISMS REGULATING FIBRONECTIN DEPOSITION

细胞外基质的组装和降解是一个动态过程 在伤口愈合、胚胎发育和转移过程中上调的基因。 纤维连接蛋白在细胞外基质中的沉积是细胞介导的 严格控制以确保基质沉积受控的工艺。 尽管导致纤维连接蛋白沉积的几个早期步骤 已经确定，导致纤维连接蛋白积聚的机制 转化为二硫化物稳定的多聚体的情况在很大程度上是未知的。我们有 鉴定了纤维连接蛋白中的二硫键异构酶活性，并提出了 为了确定这种活性是否催化了 纤维连接蛋白进入细胞外基质。研究考察了 纤维连接蛋白在细胞外基质中的沉积已被 调控细胞增殖也很难评估，因为大多数 贴壁细胞持续产生纤维连接蛋白基质。我们最近 分离的胚胎纤维连接蛋白零细胞并使其适应培养 在无血清条件下。这些细胞为我们提供了独特的 选择性诱导纤维连接蛋白聚合的机会 准确判断纤维粘连蛋白聚合的关系 和细胞功能。我们的数据表明，纤维连接蛋白增加了粘附性- 依赖生长，这种影响是由于纤维连接蛋白的过程 矩阵组件。这些研究将提供至关重要的信息 了解纤维连接蛋白在纤维连接蛋白沉积中的作用 在动脉粥样硬化和纤维化期间。申请人的即时研究 目标是：1)确定是否发生纤维连接蛋白沉积 在动脉粥样硬化和纤维化期间。申请人的即时研究 目标是：1)确定纤维连接蛋白的二硫键异构酶 活性催化纤维粘连蛋白原纤维的交联 细胞外基质；2)确定纤维连接蛋白的作用机制 基质组装正向调节细胞生长；以及3)比较 细胞生长受纤维连接蛋白基质组装和其他 细胞外基质蛋白。这些学习将促进我的长期发展 研究目标是了解细胞与其细胞之间的相互作用 细胞外环境控制细胞行为的各个方面 包括细胞的生长和迁移，以及细胞如何改变其 调控的细胞外环境的组装和拆卸 细胞外基质。这笔赠款将提供资源，以充分 利用我们新分离的纤维连接蛋白零细胞系，开发 功能阻断重组抗体的分离技术 纤维连接蛋白的保守区，并发展研究方法 纤维连接蛋白引起的细胞内信号转导事件 证词。

Assembly and degradation of extracellular matrices are dynamic processes that are up-regulated during wound healing, embryogenesis, and metastasis. Deposition of fibronectin into the extracellular matrix is a cell-mediated process that is tightly regulated to ensured controlled matrix deposition. Although several of the early steps leading to fibronectin deposition have been identified, the mechanisms leading to the accumulation of fibronectin into disulfide-stabilized multimers are largely unknown. We have identified a disulfide isomerase activity within fibronectin and propose to determine whether this activity catalyzes the crosslinking of fibronectin into the extracellular matrix. Studies examining the deposition of fibronectin into the extracellular matrix have been regulating cell proliferation has also been difficult to assess since most adherent cells continuously produce a fibronectin matrix. We recently isolated embryonic fibronectin-null cells and have adapted them to culture under serum-free conditions. These cells provide us with the unique opportunity to selectively induce fibronectin polymerization in order to precisely determine the relationship between fibronectin polymerization and cell function. Our data indicate that fibronectin increases adhesion- dependent growth and that this effect is due to the process of fibronectin matrix assembly. These studies will provide information crucial to understanding the role of fibronectin fibronectin deposition, as occurs during atherosclerosis and fibrosis. The applicant's immediate research goals are: 1) to determine whether fibronectin deposition, as occurs during atherosclerosis and fibrosis. The applicant's immediate research goals are: 1) to determine whether fibronectin's disulfide isomerase activity catalyzes the crosslinking of fibronectin fibrils in the extracellular matrix; 2) to determine the mechanisms by which fibronectin matrix assembly positively regulates cell growth; and 3) to compare how cell growth is regulated by fibronectin matrix assembly and other extracellular matrix proteins. These studies will further my long term research goals of understanding how interactions of cells with their extracellular environment control various aspects of cell behavior including cell growth and migration, and how cells alter their extracellular environment by the regulated assembly and disassembly of extracellular matrices. This grant will provide the resources to fully exploit our newly isolated fibronectin null cell lines, to develop the technology to isolate function blocking recombinant antibodies to conserved regions of fibronectin, and to develop methods to study intracellular signaling events that are a consequence of fibronectin deposition.