High sensitivity LC-MS to understand the role of Proteomes in the rules of life for Plant scientists and N8 partners

高灵敏度 LC-MS 可帮助植物科学家和 N8 合作伙伴了解蛋白质组在生命规则中的作用

• 批准号: BB/W019825/1
• 负责人: Ari Sadanandom
• 金额: $ 53.8万
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019825%2F1
• 关键词: sensitivity; LC; MS; understand; role

It has become apparent that biological complexity is largely orchestrated not through the number of genes but through variation at the protein level. Post-translational modifications (PTMs) generate different forms of a protein called proteoforms. These are responsible for cell signaling in almost every biological process. The dynamic nature of PTMs allows organisms to respond to highly transient changes in the environment with precise fine-tuning. This is especially relevant to plants due to their sessile nature; their growth and development is dependent upon their ability to adapt to changes in the environment. The reliance upon PTMs for plant responses to environmental change is orchestrated by thousands of genes encoding components of PTM systems and expansion in genes encoding the PTM machinery in plants is more significant than in any other kingdom. Understanding how these proteins affect cellular responses to environmental stress is a critical component of plant breeding programmes. This will be central in addressing the global challenge of sustainably producing food in a changing climate.To date, a number of genetic variants which alter the PTM state of the corresponding protein have been found to be extremely valuable in agriculture. Examples are found in the DELLA genes, which result in a high-yielding variety of wheat that, due to the reduced height, is more resistant to damage from wind and rain. This valuable trait is controlled by a single mutant dwarfing DELLA, the rht-1 allele. The wheat rht1 allele produces a DELLA protein that is altered in its ubiquitinated state and acts as a dominant allele to increase resistance to adverse environmental stress, indicating that PTMs can have added value in expediting breeding programmes with dominant alleles. It is predicted that the critical difference between the flooding tolerant and intolerant SUB1A alleles in rice is the mutation of a phosphorylation site in the SUB1A protein, further highlighting the importance of PTMs in crop adaptation to the environment. Introgression of the flooding tolerant SUB1 allele is the single most significant advancement for generating flood-tolerant rice varieties in the last 30 years. PTMs can thus be exploited to generate novel alleles for boosting crop productivity. However, a systematic approach to exploit PTMs for plant improvement strategies has been limited by the lack of appropriate methodologies for target discovery, mass spectrometry machine access and training. With our track record in PTM analysis, especially in plants, we are proposing to bridge this gap by building a plant-cell-focused proteome research platform that will be used for method development and discovery of novel PTMs in crop species. This will add significantly to the huge tapestry of genomics data held by the UK and international model plant and crop communities and provide data for the design and implementation of future research and breeding programmes.The acquisition of a highly sensitive mass spectrometer, such as the Bruker timsTOF Pro 2, with capability to detect these often-ephemeral modifications (as detailed in the proposal), will be central to our goal. The spectrometer will identify PTMs associated with beneficial traits while other technologies will elucidate the cellular responses to them. Although the focus is on plants the new mass spectrometer will equally serve a range of animal and microbial scientists in Durham who investigate PTM mediated signalling to understand the rules of life in various organisms. In some aspects PTM analysis in animal and microbial fields is further advanced than in plants and cross fertilisation of knowledge between these fields brings added value to this proposal.

显然，生物复杂性在很大程度上不是通过基因的数量而是通过蛋白质水平的变异来精心策划的。翻译后修饰（PTMS）产生不同形式的蛋白质形式，称为蛋白质成型。这些几乎在每个生物过程中负责细胞信号传导。 PTM的动态性质使生物体可以通过精确的微调来应对环境中的高度短暂变化。这与植物尤其相关，因为它们的无柄性质。他们的成长和发展取决于他们适应环境变化的能力。编码PTM系统组成部分的基因和编码植物中PTM机械的基因扩展的数千个基因策划了对PTMS对环境变化的植物反应的依赖，比其他任何王国都更为重要。了解这些蛋白如何影响细胞对环境压力的反应是植物育种计划的关键组成部分。这将是应对在不断变化的气候下可持续产生粮食的全球挑战的核心。到目前为止，已经发现许多改变相应蛋白质的PTM状态的遗传变异在农业中非常有价值。在DELLA基因中发现了例子，这导致多种小麦种类繁多，由于高度降低，对风和雨水的损害更具抵抗力。这个有价值的特征由单个突变体矮人Della（RHT-1等位基因）控制。小麦RHT1等位基因产生一种DELLA蛋白，该蛋白在其泛素化状态下变化，并作为增加对不良环境压力的耐药性的主要等位基因，这表明PTM可以增加具有优势等位基因的育种计划的价值。据预测，水稻中洪水耐受性和不耐受的SUB1A等位基因之间的临界差异是SUB1A蛋白中磷酸化位点的突变，进一步凸显了PTMS在作物适应环境中的重要性。在过去30年中，渗透性耐受的SUB1等位基因的渗透是产生耐洪水的水稻品种的最重要进步。因此，可以利用PTMS生成新的等位基因来提高作物生产力。但是，由于缺乏适当的目标发现方法，质谱机访问和培训的方法，一种系统的方法来利用PTM进行植物改进策略。借助我们在PTM分析中的往绩，尤其是在植物中，我们建议通过建立一个以植物为中心的蛋白质组研究平台来弥合这一差距，该平台将用于方法开发和发现农作物物种中新型PTM。这将大大增加英国和国际模型植物和作物社区的大量基因组学数据，并为未来的研究和繁殖程序的设计和实施提供数据。对高度敏感的质谱仪的获取，例如Br​​uker Timstof Pro 2（例如，能够检测这些经常是全超级人物的修改）（以及在详细范围内），这是核心的核心（以及核心）。光谱仪将识别与有益性状相关的PTM，而其他技术将阐明细胞对其的反应。尽管重点是植物，但新的质谱仪将平均服务于达勒姆的一系列动物和微生物科学家，他们研究了PTM介导的信号传导以了解各种生物的生命规则。在某些方面，动物和微生物领域的PTM分析比植物中进一步发展，这些领域之间知识的交叉受精为这一建议带来了额外的价值。