Structural insights into small molecule activation of TRPC4/5 channels

TRPC4/5 通道小分子激活的结构见解

• 批准号: BB/P020208/1
• 负责人: Robin Bon
• 金额: $ 57.52万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 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=BB%2FP020208%2F1
• 关键词: Structural; insights; into; small; molecule

The cell is the basic unit of all known living organisms, and humans consist of many different types of cells, the majority of which are highly specialised. In order to function properly, cells need to communicate with their environment and with neighbouring cells. This requires the transmission of information across the cell membrane, which separates the cellular content from the extracellular environment. One major mechanism of communication is the movement across cell membranes of ions - mainly sodium, potassium, calcium and chloride - through channel-forming proteins that are located within the membrane, so-called ion channels. Many human diseases result from abnormalities in the function of ion channels, and many successful therapeutic drugs work by activating or blocking ion channels. Our research focuses on ion channels called TRPC4 and TRPC5 channels, which are increasingly recognised as potential drug targets in a variety of diseases - including cancer, heart failure, cardiovascular and metabolic disease, epilepsy and anxiety disorders - but for which the development of activators and blockers as drugs has proven difficult. For example, we previously discovered that Englerin A, a natural product isolated from an African tree used in traditional medicine, selectively kills renal cancer cells by the potent activation of TRPC4 channels. Englerin A is also a very potent activator of TRPC5 channels. However, Englerin A is too unstable and too toxic to be used as an anti-cancer drug. In this project, we will study how Englerin A interacts with TRPC4 and TRPC5 channels . We will use a combination of experimental approaches, building on the specific expertise of the different team members. For example, we will use analogues of Englerin A that can chemically react with TRPC4/5 channels, and use mass spectrometry to identify where in the channels the reactions take place. In addition, we will use state-of-the-art electron microscopes - part of a recent £17m investment by the University of Leeds and the Wellcome Trust - to determine the three-dimensional structures of TRPC4/5 channels and their complexes with Englerin A.These results will reveal how Englerin A works on the molecular level, and how the activity of TRPC4/5 channels can be regulated by small molecules. This will enable future development of drugs that targets specific TRPC4 or TRPC5 channels, which may lead to the development of the first drugs that target these channels. We will ensure the future use of our results in the drug discovery process through our ongoing collaboration with the Lead Discovery Center of the Max Planck Society, with the aim to develop drug-like molecules for clinical trials. In addition, we will publish our results in open access publications, and make our data and materials freely available through public repositories.

该单元是所有已知的生物组织的基本单位，人类由许多不同类型的细胞组成，其中大多数是高度专业化的。为了正常运行，细胞需要与环境和相邻细胞进行通信。这需要信息在整个细胞膜中的传播，这将细胞含量与细胞外环境区分开。通信的一种主要机制是通过位于膜中所谓的离子通道内的通道形成蛋白的离子，主要是钠，钾，钙和氯化物的细胞膜运动。许多人类疾病是由离子通道功能异常引起的，许多成功的治疗药物通过激活或阻断离子通道而起作用。我们的研究重点是称为TRPC4和TRPC5通道的离子通道，这些通道越来越被认为是多种疾病中的潜在药物靶标 - 包括癌症，心力衰竭，心血管和代谢性疾病，癫痫和焦虑症 - 但由于药物的发展而导致激活剂和阻断剂的发展很难。例如，我们以前发现，从传统医学中使用的非洲树中隔离的一种天然产物Englerin A有选择地杀死肾脏癌细胞通过TRPC4通道的潜在激活。 Englerin A也是TRPC5通道的潜在激活因子。但是，Englerin A太不稳定和毒性，无法用作抗癌药。在这个项目中，我们将研究Englerin A如何与TRPC4和TRPC5通道相互作用。我们将使用实验方法的组合，这是基于不同团队成员的特定专家的基础。例如，我们将使用Englerin A的类似物，该类似物可以与TRPC4/5通道化学反应，并使用质谱法来识别在通道中发生反应的位置。此外，我们将使用最先进的电子显微镜 - 利兹大学最近进行的1,700万英镑投资的一部分 - 确定TRPC4/5渠道的三维结构及其与Englerin A的复合物的三维结构。这些结果A.这些结果将揭示Englerin在分子水平上的工作方式，以及如何通过TRPC4/5频道进行调整。这将使未来针对特定TRPC4或TRPC5通道的药物开发，这可能会导致针对这些通道的第一个药物的发展。我们将通过与Max Planck Society的铅发现中心进行的持续合作来确保未来在药物发现过程中使用结果，旨在开发类似药物的分子进行临床试验。此外，我们将在开放访问出版物中发布结果，并通过公共存储库免费提供数据和材料。

项目成果

Activation of TRPC1 Channel by Metabotropic Glutamate Receptor mGluR5 Modulates Synaptic Plasticity and Spatial Working Memory.

• DOI: 10.3389/fncel.2018.00318
• 发表时间: 2018
• 期刊: Frontiers in cellular neuroscience
• 影响因子: 5.3
• 作者: Lepannetier S;Gualdani R;Tempesta S;Schakman O;Seghers F;Kreis A;Yerna X;Slimi A;de Clippele M;Tajeddine N;Voets T;Bon RS;Beech DJ;Tissir F;Gailly P
• 通讯作者: Gailly P

Need for speed: Examining protein behaviour during cryoEM grid preparation at different timescales

对速度的需求：在不同时间尺度的冷冻电镜网格制备过程中检查蛋白质行为

• DOI: 10.1101/2020.05.14.095372
• 发表时间: 2020
• 作者: Klebl D

Need for Speed: Examining Protein Behavior during CryoEM Grid Preparation at Different Timescales.

• DOI: 10.1016/j.str.2020.07.018
• 发表时间: 2020-11-03
• 期刊: Structure (London, England : 1993)
• 作者: Klebl DP;Gravett MSC;Kontziampasis D;Wright DJ;Bon RS;Monteiro DCF;Trebbin M;Sobott F;White HD;Darrow MC;Thompson RF;Muench SP
• 通讯作者: Muench SP

Xanthine-based photoaffinity probes allow assessment of ligand engagement by TRPC5 channels

• DOI: 10.26434/chemrxiv.11890128.v1
• 发表时间: 2020-02
• 期刊: RSC Chemical Biology
• 影响因子: 4.1
• 作者: C. C. Bauer-C.;Aisling Minard;Isabelle B. Pickles;K. Simmons;Eulashini Chuntharpursat-Bon;M. Burnham

Cryo-EM structures of human TRPC5 reveal interaction of a xanthine-based TRPC1/4/5 inhibitor with a conserved lipid binding site

• DOI: 10.1101/2020.04.17.047456
• 发表时间: 2020-04
• 期刊: bioRxiv
• 作者: D. J. Wright;K. Simmons;Rachel M. Johnson;D. Beech;S. Muench;R. Bon