Assaying allosteric modulators of plant immune receptors for innovative crop protection

测定植物免疫受体的变构调节剂以实现创新作物保护

• 批准号: BB/X012050/1
• 负责人: Agnieszka Bronowska
• 金额: $ 3.19万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX012050%2F1
• 关键词: Assaying; allosteric; modulators; plant; immune

This project will address fundamental questions regarding allosteric modulation of tomato PSKR1 receptors, as we will be able to accurately quantify receptor-ligand interactions through development and validation of robust and facile to use cell-free assay, based on microscale thermophoresis (MST). MST detection enables characterisation of interactions for a wide range of biomolecules, even for challenging samples such as whole cell lysates.The compelling goal of this proposal lies in the application of a fluorophore-tagged PSKR1 receptor into MST immobilisation-free technology without a requirement of protein purification. We will transiently express tomato PSKR1 in tobacco leaves to ensure native microenvironment and post-translational modifications (e.g. glycosylations), and subsequently test, validate and optimise the assay conditions for binding peptides and small molecules alike. Through this approach, we will develop a new technology underpinning research in plant immune system applied to crop protection and enabling target-focused discovery of novel protective agents. We will establish new ways to study the function of plant PSK receptors in health and disease, and involvement of plant immune receptors in host-pathogen interactions at unprecedented level of detail. We will understand how interfering with specific allosteric binding site, identified in our pilot data, impacts interactions between PSKR1 and pathogen-derived peptides showed to be involved in the disease, and how these sites can be utilised in boosting plant immune responses against the pathogens. Technology and proof-of-concept data generated in this proposal will be used to support the larger funding bid that we plan with our project partner and collaborators, and will also provide a route for commercialisation through target validation of PSKR1 and discovery of novel chemical entities that will be jointly developed into novel crop protective agents.Proposed research, spanning biomolecular interactions, biochemistry, cell, molecular and plant biology will have an impact on in the areas of crop protection, plant immunology, training of early career scientists, and cross-sectoral engagement between academics from diverse related disciplines, industrial partners, and growers. In conclusion, this multidisciplinary proposal will yield (i) the assay, quantifying interactions between PSKR1 and ligands; (ii) proof-of-concept data on host-pathogen interactions via PSKR1 receptors; (iii) development of allosteric modulator leads to enable development of new agents for crop protection, to underpin future grant applications, and to establish a new intellectual property.

该项目将解决有关番茄PSKR 1受体变构调节的基本问题，因为我们将能够通过开发和验证基于微尺度热泳（MST）的强大且易于使用的无细胞测定来准确定量受体-配体相互作用。MST检测能够表征各种生物分子的相互作用，甚至是具有挑战性的样品，如全细胞裂解物。该提案的令人信服的目标在于将荧光团标记的PSKR 1受体应用于MST免固定技术，而无需蛋白纯化。我们将在烟叶中瞬时表达番茄PSKR 1，以确保天然微环境和翻译后修饰（例如糖基化），并随后测试，验证和优化结合肽和小分子的测定条件。通过这种方法，我们将开发一种新技术，支持应用于作物保护的植物免疫系统的研究，并实现以目标为中心的新型保护剂的发现。我们将建立新的方法来研究植物PSK受体在健康和疾病中的功能，以及植物免疫受体在前所未有的细节水平上参与宿主-病原体相互作用。我们将了解如何干扰在我们的试点数据中确定的特定变构结合位点，影响PSKR 1和病原体衍生肽之间的相互作用，这些肽与疾病有关，以及如何利用这些位点来增强植物对病原体的免疫反应。该提案中产生的技术和概念验证数据将用于支持我们与项目合作伙伴和合作者计划的更大资金竞标，并将通过PSKR 1的目标验证和发现将共同开发为新型作物保护剂的新型化学实体提供商业化途径。拟议的研究，涵盖生物分子相互作用，生物化学，细胞，分子和植物生物学将对作物保护、植物免疫学、早期职业科学家培训以及来自不同相关学科的学者、工业合作伙伴和种植者之间的跨部门接触等领域产生影响。总之，这项多学科的提案将产生（i）测定，量化PSKR 1和配体之间的相互作用;（ii）通过PSKR 1受体的宿主-病原体相互作用的概念验证数据;（iii）开发变构调节剂，从而能够开发用于作物保护的新药剂，支持未来的拨款申请，并建立新的知识产权。