Design and Synthesis of New Chemical Probes and Their Applications to Understanding Protein Structure and Function by NMR

新型化学探针的设计和合成及其在通过核磁共振了解蛋白质结构和功能中的应用

• 批准号: 2289418
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2289418
• 关键词: Design; Synthesis; New; Chemical; Probes

Understanding protein structure and function, including the mechanisms of protein-protein and protein-small molecule interactions, is challenging and requires the use of a range of sophisticated techniques including X-ray crystallography, NMR and cryoEM. Recently we designed a powerful in vitro NMR assay to monitor protein-bound biosynthetic transformations using a single carbon-13 tracer in a complex assay without any need for purification.The aim of this project is to build upon this exciting foundation and expand this basic technique for wide application by the scientific community. We will focus on the elucidation of enzyme mechanisms to facilitate the development of biocatalysts for use in the clean and efficient synthesis of biological active targets (with a focus on antibiotics) and analysis of the conformation of proteins, including, for example, a-synuclein which is implicated in Parkinson's disease.This project will involve the design and synthesis of the NMR probes with selective incorporation of carbon-13 label(s). Syntheses of isotopically labelled compounds are challenging, due to the limited availability of suitably labelled starting materials and their costs. Hence efficient synthetic strategies are required. These probes will either be ligated to the protein or used as the small molecule component of the system under study. Assessment will then be conducted by high field NMR spectroscopy. The benefits of replacing protons adjacent to the carbon-13 label with deuterium to help to sharpen the NMR signals of large systems will be investigated.

了解蛋白质的结构和功能，包括蛋白质-蛋白质和蛋白质-小分子相互作用的机制，是具有挑战性的，需要使用一系列复杂的技术，包括x射线晶体学，核磁共振和低温电子显微镜。最近，我们设计了一种强大的体外核磁共振检测方法来监测蛋白质结合的生物合成转化，在复杂的检测中使用单一的碳-13示踪剂，而无需任何纯化。该项目的目的是建立在这个令人兴奋的基础上，并将这一基本技术扩展到科学界的广泛应用。我们将重点阐明酶的机制，以促进生物催化剂的发展，用于清洁和有效地合成生物活性靶点（重点是抗生素）和分析蛋白质的构象，包括，例如，与帕金森病有关的a-突触核蛋白。该项目将涉及设计和合成具有选择性碳-13标签的核磁共振探针。由于适当标记的起始材料的可用性和成本有限，同位素标记化合物的合成具有挑战性。因此，需要有效的综合策略。这些探针要么连接到蛋白质上，要么用作所研究系统的小分子成分。然后将通过高场核磁共振波谱进行评估。用氘取代碳-13标签附近的质子，以帮助增强大型系统的核磁共振信号的好处将被研究。