Dissecting the ABA signalling pathway with a biological chemistry approach

用生物化学方法剖析 ABA 信号通路

• 批准号: 1916062
• 金额: --
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1916062
• 关键词: Dissecting; ABA; signalling; pathway; biological

Abscisic acid (ABA) is a key plant hormone that controls a wide range of processes from plant development to biotic and abiotic stress responses as plants synthesize ABA to initiate various defense mechanisms. One family of key players in this process are the ABA receptors that are members of the so-called pyrabactin resistance (PYR) and PYR-like (PYL) proteins. This diverse family of PYR/PYL proteins consists of at least 14 members in Arabidopsis that all function in ABA binding and consequent signalling. While previous structural studies of the PYL1 protein have revealed a well-conserved ABA binding pocket and a binding site for members of the 2C protein phosphatases (PP2C) responsible for signal transduction, the diverse roles of the PYL1 proteins in development are poorly understood.Aim: We have identified a series of chemical compounds with distinct biological effects on plant growth and development that are thought to be PYL1 ligands. These compounds can be used as chemical probes to further elucidate the roles of different family members in stress response and signal transduction. Furthermore, the compounds possess the potential to control germination and development, an area of great interest to our industrial partner, Tozer Seeds, Lt. The aim of this project is to investigate the molecular basis of ligand binding using a combination of biophysical and structural methods to design and synthesize chemical compounds with improved biological activities.Methods: Protein targets will be expressed in E. coli - based well-established methods using synthetic gene constructs. Biophysical methods range from high-throughput thermal shift assays (TSA) to highly accurate isothermal titration calorimetry (ITC). Co-crystallisation experiment will begin with the known condition for PYL1 and will then be extended to other family members. Structural and biophysical data will be used to guide the synthesis of new compounds. This project represents a collaboration with Tozer Seeds Ltd, UK's largest independent vegetable seed company,where biological tests will be performed during a 3-month placement.

脱落酸（阿坝）是一种重要的植物激素，在植物合成阿坝启动各种防御机制的过程中，控制着从植物发育到生物和非生物胁迫反应的广泛过程。这个过程中的一个关键参与者家族是阿坝受体，它们是所谓的吡拉菌素抗性（PYR）和PYR样（PYL）蛋白的成员。PYR/PYL蛋白的这个不同的家族由拟南芥中的至少14个成员组成，所有成员都在阿坝结合和随后的信号传导中起作用。虽然以前的结构研究的PYL 1蛋白揭示了一个保守的阿坝结合口袋和一个结合位点的成员的2C蛋白磷酸酶（PP 2C）负责信号转导，不同的角色的PYL 1蛋白在发展中知之甚少。目的：我们已经确定了一系列的化学化合物与不同的生物学效应对植物的生长和发育，被认为是PYL 1配体。这些化合物可作为化学探针，进一步阐明不同家族成员在应激反应和信号转导中的作用。此外，化合物具有控制发芽和发展的潜力，我们的工业合作伙伴，Tozer Seeds，Lt.该项目的目的是研究配体结合的分子基础，使用生物物理和结构的方法相结合，设计和合成具有改善的生物活性的化合物。基于大肠杆菌的成熟方法使用合成基因构建体。生物物理方法的范围从高通量热位移测定（TSA）到高精度等温滴定量热法（ITC）。共结晶实验将从PYL 1的已知条件开始，然后扩展到其他家族成员。结构和生物物理数据将用于指导新化合物的合成。该项目代表了与英国最大的独立蔬菜种子公司Tozer Seeds Ltd的合作，将在3个月的放置期间进行生物测试。