Structural, biochemical and genetic analysis of the newly discovered Apaf-1-binding protein CABY

新发现的Apaf-1结合蛋白CABY的结构、生化和遗传分析

• 批准号: 114626271
• 负责人: Professor Dr. Volker Dötsch
• 金额: --
• 依托单位: Else Kröner-Fresenius-Stiftung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/114626271?language=en
• 关键词: Structural; biochemical; genetic; analysis; newly

Inhibition of apoptosis is recognized as a major cause of tumorigenesis, and identification of further apoptosis-regulating proteins as potential targets for molecular therapy has become an important goal. During activation of the mitochondrial apoptosis pathway, the large cytosolic apoptosome complex is assembled, which consists of the oligomerized adaptor protein Apaf-1, Cytochrome c, dATP/ATP and Caspase-9. Apoptosome formation leads to the further recruitment and activation of the effector Caspase-3 and, subsequently, to cell death. This pathway is activated by a plethora of stimuli, including radiation and chemotherapy during cancer treatment. We recently discovered a novel Apaf-1-binding protein, which we named CABY and which sensitizes cells to death-inducing stimuli activating the mitochondrial apoptosis pathway. In the proposed project we plan to carry out a detailed molecular and structural analysis of CABY. For this purpose, we will combine biochemical approaches with NMR spectroscopy to uncover the precise molecular mechanism through which CABY influences activation of the apoptosome complex. Genetic studies using a conditional CABY knockout mouse model will allow us to investigate the physiological function of CABY. Pro-apoptotic proteins may function as tumor suppressor molecules. Our previous results have shown that CABY is downregulated in gastrointestinal stromal tumors (GISTs), and we will perform expression studies to find out whether CABY activity is diminished in other tumor types. Collectively, the experiments will contribute to an improved understanding of apoptosis regulation and the development of new therapeutic approaches for cancer and cancer resistance.

抑制细胞凋亡被认为是肿瘤发生的主要原因，并且鉴定出进一步的细胞凋亡调节蛋白作为分子疗法的潜在靶标已成为重要目标。在激活线粒体凋亡途径期间，组装了大的胞质凋亡组复合物，由寡聚的衔接蛋白APAF-1，细胞色素C，Datp/ATP/ATP和Caspase-9组成。凋亡组的形成导致效应子caspase-3的进一步募集和激活，然后导致细胞死亡。该途径被大量刺激激活，包括癌症治疗期间的放射和化学疗法。我们最近发现了一种新型的APAF-1结合蛋白，我们将其命名为Caby，并将细胞敏感到激活线粒体凋亡途径的诱导刺激。在拟议的项目中，我们计划对Caby进行详细的分子和结构分析。为此，我们将将生化方法与NMR光谱结合起来，以揭示Caby会影响凋亡组复合物激活的精确分子机制。使用有条件的Caby敲除小鼠模型的遗传研究将使我们能够研究Caby的生理功能。促凋亡蛋白可以用作肿瘤抑制分子。我们先前的结果表明，在胃肠道肿瘤（GIST）中，Caby被下调，我们将进行表达研究，以找出其他肿瘤类型中的Caby活性是否减少。总的来说，这些实验将有助于改善对凋亡调节的理解，并开发用于癌症和癌症抗癌的新治疗方法。