Conformational Disorder in Protein Function and Pathogenesis

执行摘要：我的研究计划的总体目标是揭示驱动蛋白质生物活性-有时是病理学-的分子机制。我的团队的方法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，因为它们介导了在分子水平上定义生命的过程。目前的建议建立在我们先前的成功基础上，能够更深入地探索：（i）酶催化中的构象动力学（ii）内在无序蛋白质中的功能和病理学（iii）蛋白质中活性调节的结构和动力学基础（iv）用于定量测量蛋白质相互作用的氢/氘交换背景：结构生物学的经典工具（即，X射线晶体学和经典的结构核磁共振（NMR）提供了蛋白质“天然结构”的精致详细的“快照”。然而，如果所有的蛋白质实际上都像它们在这些“快照”中出现的那样静止，那么大多数蛋白质将完全没有功能。为了实现生物活性，蛋白质必须在“类天然”结构的集合中获得特定的更高能量的构象，这些结构通过称为“构象动力学”的热驱动波动来填充。我的团队开发了一套独特的基于质谱的工具，可以提供构象动力学的详细图像，以及几乎所有生物过程的基础构象变化。在此基础上，我们探索了驱动蛋白质催化，络合，蛋白质折叠和活性调节的分子过程。影响：拟议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。这是雄心勃勃的，但直接建立在我前两个发现时期的基础上，我的团队在传统学术标准下产生了巨大的影响（在高影响力期刊上发表了53篇文章，邀请了70多场演讲等）。我们的技术已被加拿大、美国、中国、以色列、新西兰、瑞典和英国的集团复制。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。正是因为这一成功，我们更广泛的研究计划现在包括不同的，但严重交织的分支发现和知识翻译/动员。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本探索对“真实的世界”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。

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.