Mechanisms of transmembrane signalling by tetraspanins

四跨膜蛋白跨膜信号传导机制

• 批准号: G0601073/1
• 负责人: Michael Overduin
• 金额: $ 59.08万
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=G0601073%2F1
• 关键词: Mechanisms; transmembrane; signalling; tetraspanins

In this project the structures and interactions of the human tetraspanin proteins will be investigated by heteronuclear magnetic resonance spectroscopy. We have discovered that tetraspanin C-termini are bound by the tandem PDZ proteins of syntenin, and intend to elucidate the structural basis of these interactions in order to understand how they influence receptor signaling and endocytosis. The novel regulatory effects of PDZ domain phosphorylation and stabilizing terminal extensions will be investigated in order to build a model of the signaling mechanism. The extracellular domain has been expressed for NMR analysis of its lipid interaction sites and protein docking studies, providing a basis for defining its function as an cell surface receptor. Tetraspanins have been expressed as functionally intact full length proteins, and production will be scaled up to analyse their structural and binding properties in vitro. In particular, we seek to determine the oligomeric state and structural properties of the proteins in the free and ligand bound states using mixed micelles to solubilize the intact receptor. Together with collaborative in vivo cell biological studies this will provide a better understanding of the structural mechanism of tetraspanin signaling.

在该项目中，将通过异核磁共振波谱研究人类四跨膜蛋白的结构和相互作用。我们发现四跨膜蛋白 C 末端与 Syntenin 的串联 PDZ 蛋白结合，并打算阐明这些相互作用的结构基础，以了解它们如何影响受体信号传导和内吞作用。将研究 PDZ 结构域磷酸化和稳定末端延伸的新颖调节作用，以建立信号传导机制的模型。细胞外结构域已被表达，用于其脂质相互作用位点的核磁共振分析和蛋白质对接研究，为定义其作为细胞表面受体的功能提供了基础。四跨膜蛋白已被表达为功能完整的全长蛋白质，并且将扩大生产规模以分析其体外结构和结合特性。特别是，我们寻求使用混合胶束溶解完整受体来确定游离状态和配体结合状态的蛋白质的寡聚状态和结构特性。与体内细胞生物学合作研究一起，这将有助于更好地理解四跨膜蛋白信号传导的结构机制。