Functional Characterisation of insect nicotinic Acetylcholine Receptors

昆虫烟碱乙酰胆碱受体的功能表征

• 批准号: BB/P021107/1
• 负责人: Kathryn Lilley
• 金额: $ 82.94万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP021107%2F1
• 关键词: Functional; Characterisation; insect; nicotinic; Acetylcholine

By 2050 it is estimated that the global population will have increased by a third. The calorific requirements to sustain this population will need to increase by 50% and, with a growing preference for meat consumption, additional provision of feedstocks will also be needed. It has been estimated that this will require a 70% increase in crop yields by 2050. Thus increasing demands will be placed on available agricultural land to deliver efficient, reliable and sustainable food production. Insecticides are important tools for ensuring efficient crop yields. New insecticides need to be created on a regular basis to overcome resistance in pest populations as well as providing more benign environmental impacts and human safety profiles. In particular, new insecticides need to exhibit low toxicity to non-target species, particularly the major pollinators such as bees, while retaining efficacy against both the 'chewing' and 'sucking' pests that can devastate many major crops. The target of the most commercially important insecticides is the neuronal acetylcholine signalling pathway and of this class of insecticides, the safest and most effective are molecules (neonicotinoids and spinosyns) that act at the nicotinic acetylcholine receptor (nAChR). The nAChR is in fact composed of five protein subunits that form a complex. In the well established model insect, the fruit fly Drosophila melanogaster, there are ten different genes encoding nAChR subunit proteins, and these can combine to form many different classes of nAChRs, each with the potential to interact differently with insecticidal compounds. To date, very little is known about how these subunits combine, what other proteins act as accessory molecules interacting with receptor subunits, or what impact the 'parts list' of each complex has on its pharmacological properties. The aim of this project is to characterise functionally distinct classes of nAChRs in Drosophila melanogaster, a genetically and biochemically tractable insect model system that will generate insights broadly applicable to other insect species. Knowledge of the functional classes of nAChRs in Drosophila will pave the way for more targeted future research in the pest insects that cause havoc to major crops and for which there are currently very few genetics tools. In this application we will apply cutting edge spatial proteomics methods created by Kathryn Lilley with insect gene editing and advanced genetics techniques available from Steve Russell to advance the insect receptor biochemical and pharmacological approaches established at Syngenta, one of the world's leading agrochemical companies. Together, the proposed research programme will lead to a thorough characterisation of different native nAChR classes and substantially advance our understanding of a class of insecticide targets that are crucial for protecting global agriculture. The ability to design effective new insecticides that are safe and have low environmental impact will be essential to the continuing drive to increase agricultural yields and better feed the growing global population.

据估计，到2050年，全球人口将增加三分之一。维持这一人口的热量需求将需要增加50%，随着对肉类消费的日益偏好，还需要额外提供饲料。据估计，到2050年，这将需要作物产量增加70%。因此，对可用农业土地的需求将日益增加，以提供高效、可靠和可持续的粮食生产。杀虫剂是确保作物高产的重要工具。需要定期开发新的杀虫剂，以克服害虫种群的抗药性，并提供更良性的环境影响和人类安全状况。特别是，新的杀虫剂需要对非目标物种表现出低毒性，特别是主要的授粉者，如蜜蜂，同时保持对可能危害许多主要作物的“咀嚼”和“吸吮”害虫的功效。商业上最重要的杀虫剂的靶标是神经元乙酰胆碱信号传导途径，并且在这类杀虫剂中，最安全和最有效的是作用于烟碱乙酰胆碱受体（nAChR）的分子（类烟碱和多杀菌素）。nAChR实际上由五个蛋白质亚基组成，形成一个复合物。在已建立的模式昆虫果蝇（Drosophila melanogaster）中，有10种不同的编码nAChR亚基蛋白的基因，这些基因可以联合收割机形成许多不同类别的nAChR，每种nAChR都具有与杀虫化合物不同地相互作用的潜力。到目前为止，人们对这些亚基如何联合收割机，还有哪些蛋白质作为辅助分子与受体亚基相互作用，或者每个复合物的“部件清单”对其药理学性质有什么影响知之甚少。该项目的目的是研究果蝇中功能不同的nAChR类，果蝇是一种遗传和生物化学上易于处理的昆虫模型系统，将产生广泛适用于其他昆虫物种的见解。了解果蝇中nAChRs的功能类别将为未来更有针对性地研究对主要作物造成严重破坏的害虫铺平道路，目前很少有遗传学工具。在本申请中，我们将应用Kathryn Lilley创建的尖端空间蛋白质组学方法，以及Steve Russell提供的昆虫基因编辑和先进遗传学技术，以推进世界领先的农业化学公司之一先正达建立的昆虫受体生化和药理学方法。总之，拟议的研究计划将导致不同的本地nAChR类的彻底表征，并大大推进我们对一类杀虫剂目标的理解，这对保护全球农业至关重要。设计安全且对环境影响小的有效新杀虫剂的能力，对于继续努力提高农业产量和更好地养活不断增长的全球人口至关重要。

项目成果

Drosophila nicotinic acetylcholine receptor subunits and their native interactions with insecticidal peptide toxins

• DOI: 10.1101/2021.08.13.456240
• 发表时间: 2021-08
• 期刊: eLife
• 影响因子: 7.7
• 作者: D. Korona;Benedict Dirnberger;Carlo N G Giachello;R. Queiroz;David P. Minde;M. Deery;G. Johnson;Karin H. Müller;L. Firth;F. Earley;S. Russell;K. Lilley

Drosophila nicotinic acetylcholine receptor subunits and their native interactions with insecticidal peptide toxins.

• DOI: 10.7554/elife.74322
• 发表时间: 2022-05-16
• 期刊: ELIFE
• 影响因子: 7.7
• 作者: Korona, Dagmara;Dirnberger, Benedict;Giachello, Carlo N. G.;Queiroz, Rayner M. L.;Popovic, Rebeka;Mueller, Karin H.;Minde, David-Paul;Deery, Michael J.;Johnson, Glynnis;Firth, Lucy C.;Earley, Fergus G.;Russell, Steven;Lilley, Kathryn S.;Pless, Stephan A.
• 通讯作者: Pless, Stephan A.

Enrichment of Membrane Proteins for Downstream Analysis Using Styrene Maleic Acid Lipid Particles (SMALPs) Extraction.

• DOI: 10.21769/bioprotoc.4728
• 发表时间: 2023-08-05
• 期刊: Bio-protocol
• 影响因子: 0.8