Conformational Disorder in Protein Function and Pathogenesis

蛋白质功能和发病机制中的构象紊乱

• 批准号: RGPIN-2019-06696
• 负责人: Wilson, Derek
• 金额: $ 2.11万
• 依托单位: York University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=683036
• 关键词: Conformational; Disorder; Protein; Function; Pathogenesis

EXECUTIVE SUMMARY: The overarching objective of my research program is to uncover the molecular mechanisms that drive biological activity - and sometimes pathology - in proteins. My group's approach employs unique technologies developed in-house that allow us to monitor 'active' proteins directly as they mediate the processes that define life at the molecular level. The current proposal builds on our prior successes, enabling a deeper exploration of: ***(i) Conformational dynamics in enzyme catalysis ***(ii) Function and pathology in intrinsically disordered proteins***(iii) The structural and dynamic basis of activity modulation in proteins***(iv) Hydrogen / deuterium exchange for quantitative measurements of protein interactions ******BACKGROUND: The classical tools of structural biology (i.e., X-ray crystallography and classical structural NMR) provide exquisitely detailed 'snapshots' of the 'native structure' of proteins. However, if all proteins were actually as static as they appear in these 'snapshots', most would be completely non-functional. To achieve biological activity, proteins must access specific, higher energy conformations within an ensemble of 'native-like' structures that are populated via thermally-driven fluctuations known as 'conformational dynamics'. My group has developed a unique set of mass spectrometry-based tools that provide a detailed picture of conformational dynamics, and the conformational shifts that underlie virtually all biological processes. From this foundation, we explore the molecular processes that drive catalysis, complexation, protein folding and activity modulation in proteins.******IMPACT: The proposed research addresses fundamental questions that are of great significance to our understanding of protein function and pathogenesis at the molecular level. It is ambitious, but builds directly upon the foundation laid out in my previous two discovery periods, where my group has had substantial impact as measured by conventional academic standards (53 publications in high impact journals, over 70 invited talks etc.). Our technologies have been replicated by groups in Canada, the US, China, Isreal, New Zealand, Sweden and the UK. We have also had an unusually high degree of success at translating fundamental discoveries to practical applications in collaboration with numerous industrial partners. It is because of this success that our broader research program now includes distinct but heavily intertwined branches for discovery and knowledge translation / mobilization. These branches exhibit positive feedback, allowing us now to implement a discovery research program where our fundamental explorations of molecular mechanisms have a tangible impact on 'real world' applications, Canadian business trajectories and, ultimately, greatly enhanced training and career opportunities for HQP.*****

摘要：我的研究计划的总体目标是揭示驱动生物活性的分子机制-有时是病理-在蛋白质中。我的团队采用了内部开发的独特技术，使我们能够直接监测“活性”蛋白质，因为它们在分子水平上调节定义生命的过程。目前的建议建立在我们之前的成功基础上，使我们能够更深入地探索：***(i)酶催化的构象动力学***(ii)内在无序蛋白质的功能和病理***(iii)蛋白质活性调节的结构和动态基础***(iv)用于定量测量蛋白质相互作用的氢/氘交换******结构生物学的经典工具（即x射线晶体学和经典结构核磁共振）提供了蛋白质“天然结构”的精致详细的“快照”。然而，如果所有的蛋白质都像这些“快照”中显示的那样是静态的，那么大多数蛋白质将完全没有功能。为了获得生物活性，蛋白质必须在“天然”结构的集合中获得特定的、更高能量的构象，这些结构是通过被称为“构象动力学”的热驱动波动填充的。我的团队开发了一套独特的基于质谱的工具，提供了构象动力学的详细图像，以及几乎所有生物过程背后的构象变化。从这个基础上，我们探索驱动催化，络合，蛋白质折叠和蛋白质活性调节的分子过程。******影响：提出的研究解决了对我们在分子水平上理解蛋白质功能和发病机制具有重要意义的基本问题。它是雄心勃勃的，但直接建立在我前两个发现时期奠定的基础上，在这两个时期，我的团队以传统的学术标准衡量具有重大影响（在高影响力期刊上发表了53篇论文，70多篇受邀演讲等）。我们的技术已被加拿大、美国、中国、以色列、新西兰、瑞典和英国的团体复制。我们还与众多工业伙伴合作，在将基础发现转化为实际应用方面取得了不同寻常的高度成功。正是由于这一成功，我们更广泛的研究项目现在包括了发现和知识转化/动员的不同但紧密交织的分支。这些分支机构表现出积极的反馈，使我们现在能够实施一项发现研究计划，我们对分子机制的基础探索对“现实世界”的应用、加拿大的商业轨迹产生了切实的影响，并最终大大提高了HQP.*****的培训和职业机会