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、胚胎发生和转移过程中上调。 纤连蛋白沉积到细胞外基质中是细胞介导的 严格控制的工艺，以确保受控的基质沉积。 虽然导致纤连蛋白沉积的几个早期步骤已经被证实是一个重要的过程， 已经确定，导致纤维连接蛋白积聚的机制 转化为二硫键稳定的多聚体的方法在很大程度上是未知的。我们有 鉴定了纤连蛋白内的二硫键异构酶活性，并提出 为了确定该活性是否催化交联 纤连蛋白进入细胞外基质。研究审查 纤连蛋白沉积到细胞外基质中， 调节细胞增殖也很难评估， 贴壁细胞连续产生纤连蛋白基质。我们最近 分离的胚胎纤连蛋白缺失细胞，并使其适应培养 在无血清条件下。这些细胞为我们提供了 有机会选择性地诱导纤连蛋白聚合， 精确确定纤维连接蛋白聚合之间的关系 和细胞功能。我们的数据表明纤维连接蛋白增加了粘附- 依赖性生长，这种效应是由于纤维连接蛋白的过程 矩阵装配这些研究将提供重要信息， 了解纤维连接蛋白的作用纤维连接蛋白沉积， 动脉粥样硬化和纤维化。申请人的直接研究 目的是：1）确定纤维连接蛋白是否沉积，如发生 动脉粥样硬化和纤维化。申请人的直接研究 目的是：1）确定纤连蛋白的二硫键异构酶是否 活性催化纤维连接蛋白原纤维在 细胞外基质; 2）确定纤连蛋白 基质组装正调控细胞生长;和3）比较如何 细胞生长受纤连蛋白基质组装和其他细胞因子的调节。 细胞外基质蛋白这些研究将促进我的长期 研究目标是了解细胞与它们的 细胞外环境控制细胞行为的各个方面 包括细胞生长和迁移，以及细胞如何改变它们的 细胞外环境的调节组装和拆卸 细胞外基质这笔赠款将提供资源， 利用我们新分离的纤连蛋白空细胞系， 分离功能阻断性重组抗体的技术 纤维连接蛋白的保守区域，并开发研究方法 细胞内信号事件是纤连蛋白的结果 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.