Coiled-coil Technology for Regulating Intracellular Protein-protein Interactions

用于调节细胞内蛋白质-蛋白质相互作用的卷曲螺旋技术

• 批准号: BB/V008412/2
• 负责人: Andrew Wilson
• 金额: $ 25.01万
• 依托单位: University of Birmingham
• 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: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV008412%2F2
• 关键词: Coiled; coil; Technology; Regulating; Intracellular

Proteins are the workhorses of biology. Proteins rarely work alone, and they cooperative via so called protein-protein interactions. In this way, they form larger assemblies and networks of proteins. In turn, these provide frameworks for controlling all cellular processes that regulate life. Dysregulation of the underlying protein-protein interactions can result in disease. The scale of this framework of protein-protein interactions within a cell is enormous; it has been estimated to be around 650,000 different interactions. These present considerable opportunities for intervening in biological processes (a) to understand healthy cells better, and (b) to develop new therapeutics when subcellular mechanisms go wrong. There are different ways to do this. We propose a new approach that employs synthetic protein modules (i) to disrupt protein-protein interactions and (ii) to hijack endogenous cell machineries. Synthetic chemical probes-such as small-molecule drugs-function by binding to a protein target within the body. This can be used to interfere with the target protein's function to help understand its biological role and as a starting point for drug discovery. Most chemical probes bind to well-defined pockets in proteins; this is analogous to a key fitting into a lock. By contrast, the design of probes to interfere with protein-protein interactions generally requires a fundamentally different type of association between the probe and one of the interacting target proteins; analogous to a hand gripping a ball. Thus, the development of effective probes that target protein-protein interactions raises new challenges that need to be met in future chemical biology and drug discovery. Two emerging approaches are promising for this, and we propose to combine them in this grant application.The first is a synthetic-biology approach. This uses synthetic proteins called de novo coiled coils as scaffolds for building new protein-protein interactions from scratch. This is attractive because natural coiled-coil proteins exhibit an array of protein-recognition properties and we can design de novo coiled-coils with diverse structures thereby expanding their potential. The second involves the targeted destruction of cellular proteins; in essence, this is a search-and-destroy strategy that co-opts the cell's own waste-disposal machineries so as to block protein function or remove harmful proteins that cause disease. The proposed research will develop new methodologies (i) to disrupt specified protein-protein interactions and (ii) to target certain proteins of interest for degradation. In this way, we will regulate specified processes in cells. To do this, we will not use conventional small molecules as the probes for intervening in the underlying protein-protein interactions. Rather, we will employ the synthetic coiled coils and adapt these to recognise the target proteins. In addition, for the second application, the coiled coils will be modified further to link the target protein to the cell's degradation pathways. Our aim is to deliver methods and reagents that will be of use to others in studying biological function and for developing new drugs to treat disease.This work is necessarily interdisciplinary. Therefore, we bring together a team of computational and experimental chemists, biochemists and cell biologists to tackle it, and we partner with a biotech company to translate the work in timely and relevant manner.

蛋白质是生物学的主力。蛋白质很少单独工作，并且通过所谓的蛋白质 - 蛋白质相互作用进行合作。这样，它们形成了较大的组件和蛋白质网络。反过来，这些提供了控制所有调节生命的细胞过程的框架。潜在蛋白质蛋白相互作用的失调可能导致疾病。细胞内蛋白质蛋白质相互作用的框架的规模是巨大的。据估计，它约为650,000种不同的互动。这些目的是介入生物学过程的大量机会（a）更好地了解健康细胞，并且（b）当亚细胞机制出错时，可以开发新的治疗剂。有不同的方法可以做到这一点。我们提出了一种新方法，该方法采用合成蛋白模块（i）破坏蛋白质 - 蛋白质相互作用和（ii）到劫持内源性细胞机器的方法。合成化学物质通过与体内的蛋白质靶标结合而探测类似于小分子的药物功能。这可以用来干扰靶蛋白的功能，以帮助了解其生物学作用和药物发现的起点。大多数化学探针与蛋白质定义明确的口袋结合；这类似于将钥匙拟合到锁中。相比之下，干扰蛋白质 - 蛋白质相互作用的探针的设计通常需要探针与一种相互作用的靶蛋白之间的根本不同类型的关联。类似于手握住球的手。因此，靶向蛋白质蛋白质相互作用的有效探针的开发引发了在未来的化学生物学和药物发现中需要应对的新挑战。为此，有两种新兴方法有望，我们建议将它们结合在此赠款应用中。第一种是一种合成生物学方法。这使用了称为从头盘绕的卷轴的合成蛋白作为脚手架，用于从头开始构建新的蛋白质 - 蛋白质相互作用。这很有吸引力，因为天然盘绕蛋白蛋白具有一系列蛋白质识别特性，我们可以设计具有不同结构的从头盘绕螺旋，从而扩大了其潜力。第二个涉及细胞蛋白的靶向破坏。从本质上讲，这是一种搜索和破坏的策略，它可以选择细胞自身的废物含量机制，以阻止蛋白质功能或去除引起疾病的有害蛋白质。 拟议的研究将开发新的方法论（i），以破坏特定的蛋白质蛋白质相互作用，（ii）靶向某些感兴趣的蛋白质以降解。这样，我们将调节单元格中的指定过程。为此，我们不会使用常规的小分子作为探针介入下面的蛋白质蛋白质相互作用。相反，我们将采用合成的盘绕线圈并适应这些线圈以识别靶蛋白。此外，对于第二应用，将进一步修改盘绕线圈，以将靶蛋白与细胞的降解途径联系起来。我们的目的是提供在研究生物学功能和开发新药物治疗疾病的方法和试剂中的方法和试剂。这项工作必然是跨学科的。因此，我们将一个计算和实验化学家，生物化学家和细胞生物学家组成的团队汇集在一起​​，以解决它，并与一家生物技术公司合作，以及时和相关的方式翻译工作。