FUNCTIONAL STUDIES OF FOCAL ADHESION KINASE

粘着斑激酶的功能研究

• 批准号: 2022762
• 负责人: Steven K. Hanks
• 金额: $ 18.42万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2022762
• 关键词: animal tissue; biological signal transduction; cell adhesion molecules; cell growth regulation; enzyme activity; extracellular matrix; fibroblasts; genetic library; genetic techniques; genetic transcription; immunoprecipitation; integrins; phosphorylation; protein signal sequence; protein structure; protein tyrosine kinase

Cell interactions with the extracellular matrix (ECM) profoundly influence cell shape, migration, proliferation and differentiation - properties which are important for normal growth and development, and which affect many aspects of health care and disease including wound repair, angiogenesis, cancer metastasis, and atherogenesis. The mechanism by which the ECM influences cell behavior remains one of the basic unsolved problems in cell biology today. Recent evidence indicates that tyrosine- phosphorylation of proteins is a key event in the response. In support of this, a novel tyrosine kinase has been identified and shown to be localized to "focal adhesions," sites where interactions with the ECM occur via integrin receptors. Further studies indicate that this kinase, designated "Focal Adhesion Kinase" (FAK), becomes highly phosphorylated (on both tyrosine and serine sites) upon cell adhesion to fibronectin and other ECM substrates that bind to integrin receptors. These phosphorylation events are likely to be essential to an integrin-mediated signaling pathway involving FAK. The long term objective of the proposed work is to reach a full understanding of the role of FAK in bringing about ECM-influenced changes in cell behavior. Specifically, major sites of FAK phosphorylation induced upon the adherence of mouse fibroblasts to fibronectin will be determined by phosphopeptide mapping strategies, and the effect of these modifications on FAK's kinase activity, focal adhesion targeting, and ability to interact with known SH2-proteins will be evaluated. To gain further insight into FAK's signaling role, two molecular approaches will be undertaken to identify proteins that interact with FAK. One approach aims to identify novel SH2 proteins that interact with FAK phosphotyrosines. The other approach involves a powerful genetic screen for proteins that can stably interact with any part of the FAK molecule. Such proteins are likely to act as either upstream regulators or downstream effectors of FAK.

细胞与细胞外基质(ECM)的相互作用深刻影响 细胞形态、迁移、增殖和分化--特性 哪些对正常生长和发育很重要，哪些影响 医疗保健和疾病的许多方面，包括伤口修复， 血管生成、癌症转移和动脉粥样硬化。这个机制是通过 细胞外基质对细胞行为的影响仍然是一个基本悬而未决的问题 当今细胞生物学中的问题。最近的证据表明，酪氨酸- 蛋白质的磷酸化是反应中的一个关键事件。为了支持…… 这是一种新的酪氨酸激酶，已经被发现并被证明是 定位于“灶性粘连”，即与ECM相互作用的部位 通过整合素受体发生。进一步的研究表明，这种激酶， 被指定为“粘着斑激酶”(FAK)，高度磷酸化 (在酪氨酸和丝氨酸位点上)在细胞与纤维连接蛋白和 与整合素受体结合的其他ECM底物。这些 磷酸化事件很可能是整合素介导的 涉及FAK的信号通路。 拟议工作的长期目标是实现 理解FAK在ECM影响的变化中的作用 在细胞行为上。具体地说，FAK的主要磷酸化位点诱导 小鼠成纤维细胞与纤维连接蛋白的粘附性将被确定 通过磷酸肽定位策略，以及这些策略的效果 修饰FAK的激酶活性，焦点黏附靶向，以及 将评估与已知SH2-蛋白相互作用的能力。为了获得 进一步深入了解FAK的信号作用，两种分子方法将 以确定与FAK相互作用的蛋白质。一种方法 旨在鉴定与FAK相互作用的新SH2蛋白 磷酸酪氨酸。另一种方法涉及强大的基因筛查 对于能够稳定地与FAK分子的任何部分相互作用的蛋白质。 这类蛋白质可能作为上游调节因子或 FAK下游效应器。