Mechanistic characterizations of redox regulations and functions of Arf and Rho families

项目总结： 小分子GTP酶在细胞功能中起着关键作用。它们可以分为几个组，包括 Arf、Rab、Ras、Rho和Ran家族的蛋白质。ARF家族的氧化还原规则和功能 小的GTP酶是未知的。小GTP酶的Rho家族的氧化还原调节和功能仅是 部分为人所知。在不知道它们的情况下，与以下疾病相关的有效疗法的发展 它们是不切实际的。这个项目的一个长期目标是理解依赖于氧化还原的规则 以及这些蛋白质的功能。另一个目标是开发针对这些疾病的治疗干预措施。 这一目标包括鉴定新的氧化还原惰性核苷酸，这些核苷酸似乎针对氧化还原- 敏感的Arf和Rho蛋白，以阻止其氧化还原反应。 为了实现这些目标，在Arf蛋白家族中发现的先前未知的氧化还原敏感基序 将使用包括基于诱变的氧化还原的多学科方法来表征 生物化学与新的基于机理的核苷酸类似物以及质谱学 光谱学和细胞生物学方法。还将使用多学科方法来完善和 鉴定在Rho蛋白中发现的先前未知的氧化还原敏感基序。通过以下方式获得的表征 使用基于突变的氧化还原生物化学和细胞生物学以及其他分析方法将识别 新的和改进的氧化还原敏感型的前所未有的监管特征和氧化还原响应特性 在Arf和Rho GTP酶中发现的基序。这些蛋白质中这些氧化还原基序的特征将 然后根据它们新颖的氧化还原响应特性进行分类。这些分类将进一步允许 对这些氧化还原基序的详细氧化还原控制和作用的系统机制研究 小的GTP酶。然后，这一系统的机制研究将具体说明 在Arf和Rho家族的小GTP酶中发现的新的和精炼的氧化还原敏感基序的作用。 此外，这种多学科方法的氧化还原生化分析包括核苷酸的使用。 是新型氧化还原惰性核苷酸的类似物。这些类比的使用不仅仅是为了检查机制 这些氧化还原基序在小GTP酶中的作用，也为一种新的治疗方法提供了洞察力 这些基序的氧化还原脱敏可以缓解或终止与 在小的GTP酶中这些基序的氧化还原反应不受抑制。

Project Summary: Small GTPases play critical roles in cellular functions. They can be classified into several groups, including the Arf, Rab, Ras, Rho, and Ran families of proteins. The redox regulations and functions of the Arf families of small GTPases are unknown. The redox regulations and functions of the Rho families of small GTPases are only partly known. Without knowing them, the development of effective therapeutics for diseases associated with them is impractical. One long-term objective of this project is to understand the redox-dependent regulations and functions of these proteins. Another objective is to develop therapeutic interventions for these diseases. This objective includes the identification of the novel redox inert nucleotides that seem to target the redox- sensitive Arf and Rho proteins to block their redox responses. To achieve these goals, the previously unknown redox-sensitive motifs found in the Arf families of proteins will be characterized by using a multidisciplinary approach that includes mutagenesis-based redox biochemistry with novel mechanism-based nucleotide analogs as well as mass spectrometric, EPR spectroscopic, and cell biology methods. The multidisciplinary approach will also be used to refine and characterize previously unknown redox-sensitive motifs found in Rho proteins. The characterizations gained by using mutagenesis-based redox biochemistry and cell biology as well as other analytical methods will identify the unprecedented regulatory features and redox response properties of the new and refined redox-sensitive motifs found in Arf and Rho GTPases. The feature characteristics of these redox motifs in these proteins will then be classified based on their novel redox response properties. These classifications will further allow a systematic mechanistic investigation of the detailed redox control and action of these redox motifs in these small GTPases. This systematic mechanistic study will then specify the regulations of and mechanisms of the actions of the new and refined redox-sensitive motifs found in the Arf and Rho families of small GTPases. Furthermore, the redox biochemical analysis of this multidisciplinary approach includes the use of nucleotide analogs that are novel redox-inert nucleotides. The use of these analogs is not only to inspect the mechanisms of the actions of these redox motifs in the small GTPases but also to provide insight into a novel therapeutic strategy in which a redox desensitization of these motifs alleviates or terminates diseases associated with the unchecked redox response of these motifs in the small GTPases.