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.

在这个项目中，我们将用异核磁共振波谱来研究人类Tetraspanin蛋白的结构和相互作用。我们发现Tetraspanin C-末端与Syntenin的串联PDZ蛋白结合，并试图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转导和内吞作用。为了建立信号机制的模型，我们将研究PDZ结构域的磷酸化和稳定末端延伸的新的调节作用。胞外结构域已被表达，用于其脂类相互作用部位的核磁共振分析和蛋白质对接研究，为确定其作为细胞表面受体的功能提供了基础。Tetraspanins已经被表达为功能完整的全长蛋白质，并将扩大生产，以在体外分析它们的结构和结合特性。特别是，我们试图使用混合胶束来溶解完整的受体，以确定蛋白质在自由和配体结合状态下的低聚状态和结构性质。结合体内细胞生物学的合作研究，这将有助于更好地理解Tetraspanin信号的结构机制。