REGULATION OF NEUTROPHIL RECEPTOR G PROTEIN INTERACTIONS

中性粒细胞受体 G 蛋白相互作用的调节

• 批准号: 2487871
• 负责人: GARY M BOKOCH
• 金额: $ 28.86万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2487871
• 关键词: G protein; actins; antigen receptors; biological signal transduction; cell migration; chemoattractants; cytokine receptors; enzyme activity; growth factor receptors; guanosinetriphosphatases; human subject; leukocyte activation /transformation; molecular site; neutrophil; oncoprotein p21; phlebotomy; protein kinase; protein sequence; receptor binding; receptor coupling; site directed mutagenesis

DESCRIPTION: Communication between chemoattractant and growth factor receptors and downstream Rho family small GTPases initiates cellular events important for the motile responses of leukocytes and fibroblasts. Dr. Bokoch and his colleagues will investigate one such receptor-regulated pathway, the Rac/Cdc42-mediated activation of the target protein Pak (p21-activated kinase). The proposed studies address the biological pathways used by receptors to regulate the actin cytoskeleton, and will identify novel mechanisms by which Pak activity is controlled. In their first specific aim, the molecular interactions through which Pak regulates the actin cytoskeleton will be determined structurally and mechanistically in a well-defined fibroblast model system. Domains in Pak important for cytoskeletal regulation will be identified using a combination of molecular and cellular approaches. Target proteins which interact with these domains will be isolated and characterized. They will extend their studies of cytoskeletal regulation by Pak in the second aim, which will evaluate a novel GTPase-independent mechanism for Pak regulation. The molecular basis for Pak activation by membrane targeting will be investigated. The investigators will examine the significance of Pak activation by membrane association in cytoskeletal regulation of neurite development using the PC12 cell as a model system. The role of the Pak-binding Nck adapter protein in this process will also be probed. Receptors appear to control Pak activity by mechanisms distinct from direct activation via Rac or Cdc42 which include regulatory dephosphorylation reactions critical to the Pak activation process. The third aim will thus focus upon identification and purification of phosphatases that act upon Pak. Using molecular and biochemical means, they will probe the mechanisms by which phosphatases control Pak activity, and will initiate studies on the relevance of such regulatory phosphatases to receptor-mediated Pak activation.

描述：趋化剂和生长因子之间的通讯 受体和下游 Rho 家族小 GTP 酶启动细胞事件 对于白细胞和成纤维细胞的运动反应很重要。 博士。 博科赫和他的同事将研究一种这样的受体调节的 途径，Rac/Cdc42 介导的靶蛋白 Pak 激活 （p21 激活激酶）。 拟议的研究涉及生物学 受体用来调节肌动蛋白细胞骨架的途径，并将 确定控制 Pak 活性的新机制。 在他们的第一个具体目标中，Pak 所通过的分子相互作用 调节肌动蛋白细胞骨架将在结构上确定 机械地在明确的成纤维细胞模型系统中。 Pak 的域名 将使用组合来识别对细胞骨架调节重要的 分子和细胞方法。 与相互作用的靶蛋白 这些域将被隔离和表征。 他们将在以下领域扩展 Pak 细胞骨架调节的研究： 第二个目标，将评估 Pak 的新型 GTPase 独立机制 规定。 膜靶向激活 Pak 的分子基础 将被调查。 调查人员将研究其意义 神经突细胞骨架调节中膜关联的 Pak 激活 使用 PC12 单元作为模型系统进行开发。 的作用 在此过程中结合 Pak 的 Nck 接头蛋白也将被探测。 受体似乎通过不同于直接控制 Pak 活性的机制 通过 Rac 或 Cdc42 激活，其中包括调节性去磷酸化 对 Pak 激活过程至关重要的反应。 因此，第三个目标将 专注于鉴定和纯化作用于的磷酸酶 帕克.他们将利用分子和生化手段来探究其机制 磷酸酶通过其控制 Pak 活性，并将启动有关 此类调节性磷酸酶与受体介导的 Pak 的相关性 激活。