Isothermal Titration Calorimetry for Studying Macromolecular Interactions

用于研究大分子相互作用的等温滴定量热法

• 批准号: RTI-2023-00004
• 负责人: Patel, Trushar
• 金额: $ 10.93万
• 依托单位: University of Lethbridge
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748479
• 关键词: Isothermal; Titration; Calorimetry; Studying; Macromolecular

The unifying theme of our NSERC DG-funded research programs is to establish the role of biomolecular interactions (interactions of proteins with nucleic acids, small molecule interactions with proteins and nucleic acids, etc.,) in cellular processes in organisms such as viruses, bacteria, yeast, and humans. The pivotal challenge we face in advancing our research and training activities is the lack of critical infrastructure we require to study biomolecular interactions. Without this, we cannot perform interdisciplinary research to train students and researchers at the UofL. Recent advances in structural biology methods have expanded our understanding of how biomolecules communicate with each other and function. However, to study the high-resolution structures of biomolecular complexes, information on affinity and stoichiometry is essential. Therefore, we propose to install isothermal titration calorimetry (ITC). This widely used and reliable instrument aids a direct measure of the heat generated or absorbed as a result of biomolecular interaction. ITC allows the direct, rapid, and accurate determination of binding affinities and it is possible to study interactions of a wide range of unaltered biomolecules in their native state. In a single experiment, ITC accurately determines a wide range of binding constants, reaction stoichiometry, as well as thermodynamic parameters such as enthalpy and entropy. In addition to binding affinities, ITC can help elucidate the mechanisms underpinning molecular interactions. The UofL does not have an ITC instrument; hence, there is an urgent need for this critical infrastructure. In conjunction with the existing infrastructure at the UofL, the installation of ITC will accelerate HQP training in our laboratories, and in our collaborator's laboratories, and significantly advance research and training activities.

我们的NSERC DG资助的研究计划的统一主题是建立生物分子相互作用的作用（蛋白质与核酸的相互作用，小分子与蛋白质和核酸的相互作用等）在生物体如病毒、细菌、酵母和人类的细胞过程中。我们在推进研究和培训活动方面面临的关键挑战是缺乏研究生物分子相互作用所需的关键基础设施。没有这一点，我们不能进行跨学科的研究，以培养学生和研究人员在UofL。结构生物学方法的最新进展扩大了我们对生物分子如何相互通信和功能的理解。然而，为了研究生物分子复合物的高分辨率结构，亲和力和化学计量的信息是必不可少的。因此，我们建议安装等温滴定量热法（ITC）。这种广泛使用的可靠仪器有助于直接测量生物分子相互作用产生或吸收的热量。ITC允许直接、快速和准确地测定结合亲和力，并且可以研究各种未改变的生物分子在其天然状态下的相互作用。在一个单一的实验中，ITC准确地确定了广泛的结合常数，反应化学计量，以及热力学参数，如焓和熵。除了结合亲和力之外，ITC还可以帮助阐明分子相互作用的基础机制。利比里亚大学没有信息和通信技术工具，因此迫切需要这一关键基础设施。与UofL现有的基础设施相结合，ITC的安装将加速我们实验室和我们合作者实验室的HQP培训，并显着推进研究和培训活动。