MODULATION OF ALPHA 5 BETA 1 INTEGRIN

ALPHA 5 BETA 1 整合素的调节

• 批准号: 2903198
• 负责人: DAVID E BOETTIGER
• 金额: $ 27.04万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2903198
• 关键词: actins; bioengineering /biomedical engineering; biological signal transduction; biomechanics; biomedical equipment development; cell adhesion; chemical models; crosslink; fibronectins; integrins; measurement; mechanical pressure; physical model; protein isoforms; receptor binding; tissue /cell culture

DESCRIPTION (Adapted from applicant's abstract): Alpha5beta1 is a member of the integrin family of cell adhesion receptors which serve both the mechanical function of adhering cells and a signaling function in the control of many cell processes. It is likely that the mechanical and signaling aspects of alpha5beta1 function are linked since some alpha5beta1 signaling functions cannot be replaced by ligand binding alone. To examine the mechanical aspects of alpha5beta1 function and regulation of alpha5beta1 mediated cell adhesion, a spinning disc device has been engineered to provide a measurement of the force required to detach cells from a substrate. By applying concepts from chemical binding analysis it has been possible to convert the adhesion data to a model of monovalent receptor-ligand binding and demonstrate that there is a linear relationship between the number of integrin-extracellular matrix bonds and the force required to detach the cell. The proposal considers models to explain activation of alpha5beta1 binding to fibronectin: agonist activation, induced-fit activation, clustering activation, and tension activation. Each of these models makes different predictions for ligand, cytoskeletal, and energy requirements. These requirements would be tested using biochemical predictions. Inhibitors, and genetic analysis of fibronectin mutants, beta1 cytoplasmic domain splice variants, and beta1 cytoplasmic domain mutants. The analysis views the adhesion connection as one from fibronectin to alpha5beta1 to actin cytoskeleton and these connections, either ligand binding or cytoskeletal binding, can control the adhesion strength. Procedures have been developed to assay these connections separately. Their approach would be extended to the examination of long term (or steady-state) adhesion, including analysis of the increase in the number of alpha5beta1 receptors bound, and the role of clustering n the adhesion strengthening-cell spreading process.

描述（改编自申请人的摘要）：alpha5beta1是 整合素的细胞粘附受体家族，既服务于机械 粘附细胞的功能和信号传导函数在许多细胞中控制 过程。可能的机械和信令方面 由于某些alpha5beta1信号传导函数，因此链接了alpha5beta1函数 不能单独用配体结合代替。检查机械方面 alpha5beta1函数和α5BETA1介导的细胞粘附的调节，A 已经设计了旋转盘设备以提供对力的测量 需要从底物脱离细胞。通过应用化学概念 绑定分析可以将粘附数据转换为模型 单价受体 - 配体结合，并证明有线性 整联蛋白 - 托管基质键的数量与 分离细胞所需的力。该提案考虑解释模型 α5Beta1与纤连蛋白结合的激活：激动剂激活， 诱导拟合激活，聚类激活和张力激活。每个 这些模型对配体，细胞骨架和能量做出了不同的预测 要求。这些要求将使用生化预测进行测试。 抑制剂和纤连蛋白突变体的遗传分析，β1细胞质 域剪接变体和β1细胞质结构域突变体。分析 将粘附连接视为从纤连蛋白到α5Beta1再到肌动蛋白的连接 细胞骨架和这些连接，是配体结合或细胞骨架 结合，可以控制粘附强度。已经制定了程序 分别测定这些连接。他们的方法将扩展到 检查长期（或稳态）粘附，包括分析 α5BETA1受体的数量增加，以及 聚类n粘附增强细胞扩散过程。