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

项目摘要： 小GTP酶在细胞功能中起关键作用。它们可以分为几类，包括 蛋白质的Arf、Rab、Ras、Rho和Ran家族。Arf家族的氧化还原调节和功能 小GTP酶未知。小GTP酶的Rho家族的氧化还原调节和功能仅限于 部分已知。在不了解它们的情况下，开发有效的治疗方法来治疗与 他们是不切实际的。这个项目的一个长期目标是了解氧化还原依赖的调节 这些蛋白质的功能。另一个目标是为这些疾病制定治疗干预措施。 这一目标包括鉴定似乎靶向氧化还原酶的新型氧化还原惰性核苷酸。 敏感的Arf和Rho蛋白阻断它们的氧化还原反应。 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 将通过使用多学科方法来表征，包括基于诱变的氧化还原 生物化学与新的机制为基础的核苷酸类似物以及质谱，EPR 光谱和细胞生物学方法。还将采用多学科方法， 表征以前未知的氧化还原敏感性基序中发现的Rho蛋白。通过以下方式获得的表征： 使用基于诱变的氧化还原生物化学和细胞生物学以及其他分析方法， 新的和改进的氧化还原敏感的前所未有的监管功能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特性将 然后基于它们的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 详细的氧化还原控制和这些氧化还原基序在这些系统的机制调查 小GTP这种系统的机制研究，然后将详细说明的规则和机制， 在小GTP酶的Arf和Rho家族中发现的新的和精制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使用核苷酸 是新的氧化还原惰性核苷酸的类似物。使用这些类似物不仅是为了检查机制 这些氧化还原基序在小GTP酶中的作用，同时也提供了一种新的治疗方法， 其中这些基序的氧化还原脱敏可减轻或终止与这些基序相关的疾病， 这些基序在小GTP酶中的未受抑制的氧化还原反应。

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.