MODEL SYSTEM FOR STUDY OF CELL-MATRIX INTERACTION

这笔赠款的目的是开发一种可广泛应用于 细胞与细胞外基质相互作用的研究。一种新兴的 矩阵-受体领域的概念是存在固有的冗余 在整合素-配基系统中，这种受体-配基的多样性可以 提供一种机制，细胞通过该机制专门响应 细胞外基质。拟议的研究将集中于确定 这种受体-配体复制的潜在机制和提供 关于整合素调节细胞行为的重要信息- 矩阵交互作用。 作为我们的模型，我们将开发整合素α3beta1与 层粘连蛋白和纤维连接蛋白。Alpha3beta1代表多样性 与许多已知的整合素相关。具体来说，我们将 定位和描绘层粘连蛋白和纤维连接蛋白的结合部位 对于字母3beta1。此外，我们还将描述中的结合位点 层粘连蛋白和纤维连接蛋白的α3β1。这项工作将使用以下工具完成 蛋白质水解性和化学衍生片段、重组多肽和 合成肽，来源于层粘连蛋白、纤维连接蛋白和α3β1 固相结合分析、亲和层析、交联表位 用于测绘实验和生产单抗。已被占用 总之，这些研究将建立潜在的分子机制。 α3β1与层粘连蛋白和纤维连接蛋白相互作用。 我们最近演示了一种细胞类型特异性的调节 α3β1配体选择性。拟议研究的这一部分将 对我们的初步发现进行扩展，并确定可能的机制 这种细胞类型特异性地调节受体的结合。这些 将使用cdna分析和免疫学方法进行研究。 接近了。最后，拟议的研究将确定和描述 α3β1的其他配体。使用Alpha3beta1作为亲和力矩阵 已经分离出一种可能与细胞有关的新的潜在分子-细胞 通过字母3beta1结合。拟议的工作将进一步描述 这种新的糖蛋白，并探索是否有可能 α3β1相互作用的配体。 我们预计这些研究将有助于我们对 细胞选择性与基质蛋白相互作用的机制； 从而提供了对细胞行为调节的洞察力 细胞外基质。此外，对其重要意义的理解 对于这种受体-配体冗余，将从拟议的研究中进化出来。

The purpose of this grant is to develop a model with broad applications for the study of cell interaction with extracellular matrix. An emerging concept in the matrix-receptor field is that there is redundancy inherent in the integrin-ligand system and this receptor-ligand multiplicity may provide a mechanism by which cells specifically respond to the extracellular matrix. The proposed research will focus on identifying the underlying mechanisms for this receptor-ligand duplication and provide important information on the regulation of cellular behavior by integrin- matrix interactions. We will develop as our model, the binding of the integrin alpha3beta1 to laminin and fibronectin. Alpha3beta1 is representative of the multiplicity associated with many of the known integrins. Specifically, we will localized and delineate the binding sites within laminin and fibronectin for alpha3beta1. In addition, we will characterize the binding sites in alpha3beta1 for both laminin and fibronectin. This work will be done using proteolytic and chemically derived fragments, recombinant polypeptides and synthetic peptides, derived from laminin, fibronectin and alpha3beta1 in solid phase binding assays, affinity chromatography, crosslinking epitope mapping experiments and for monoclonal antibody production. Taken together, these studies will establish the molecular mechanisms underlying alpha3beta1 interaction with laminin and fibronectin. We have recently demonstrated a cell type specific modulation of alpha3beta1 ligand selectivity. This portion of the proposed research will expand on our preliminary findings and identify possible mechanisms for this cell type specific modulation of receptor binding. These investigations will be done using cDNA analysis and immunological approaches. Finally, the proposed studies will identify and characterize other ligands for alpha3beta1. Using alpha3beta1 as an affinity matrix we have isolated a new potential molecule that may be involved in cell-cell binding through alpha3beta1. The proposed work will further characterize this new glycoprotein and explore the possibilities of yet additional ligands for alpha3beta1 interaction. We anticipate that these studies will contribute to our understanding of the mechanisms by which cell selectivity interact with matrix proteins; thus providing insight into the regulation of cellular behavior by the extracellular matrix. Furthermore, an understanding of the significance for such receptor-ligand redundancy will evolve from the proposed research.