FUNCTIONAL STUDIES OF FOCAL ADHESION KINASE

粘着斑激酶的功能研究

• 批准号: 2187441
• 负责人: Steven K. Hanks
• 金额: $ 17.48万
• 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/2187441
• 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 细胞形状、迁移、增殖和分化 - 特性 这对正常生长和发育很重要，并影响 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形成。 该机制由 ECM 对细胞行为的影响仍然是未解决的基本问题之一 当今细胞生物学的问题。最近的证据表明酪氨酸- 蛋白质的磷酸化是反应中的关键事件。支持 一种新的酪氨酸激酶已被鉴定并被证明是 定位于“粘着斑”，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， 指定为“粘着斑激酶”(FAK)，变得高度磷酸化 （在酪氨酸和丝氨酸位点上）细胞粘附到纤连蛋白和 其他与整合素受体结合的 ECM 底物。这些 磷酸化事件可能对于整合素介导的 涉及FAK的信号通路。 拟议工作的长期目标是全面实现 了解 FAK 在带来 ECM 影响的变化中的作用 在细胞行为中。具体来说，FAK 磷酸化的主要位点诱导 将确定小鼠成纤维细胞对纤连蛋白的粘附 通过磷酸肽作图策略，以及这些策略的效果 对 FAK 激酶活性、粘着斑靶向性的修饰，以及 将评估与已知 SH2 蛋白相互作用的能力。为了获得 为了进一步了解 FAK 的信号传导作用，两种分子方法将 旨在鉴定与 FAK 相互作用的蛋白质。一种方法 旨在鉴定与 FAK 相互作用的新型 SH2 蛋白 磷酸酪氨酸。另一种方法涉及强大的基因筛选 对于能够与 FAK 分子的任何部分稳定相互作用的蛋白质。 这些蛋白质可能充当上游调节剂或 FAK 的下游效应器。