Protein Choreography

生命的过程是动态的--它是分子水平上的变化，使我们能够生长和移动，但也使我们生病和治疗疾病。正如我们身体的形状和姿势可以决定我们执行任务的准备程度一样，蛋白质分子的结构和构象也可以决定它的功能或活性。蛋白质的动态和快速重组能力是生物学、疾病和医学中许多关键活动的基础。然而，目前我们仅限于在空间或时间上以高分辨率研究蛋白质，包括许多重要的酶--但不是两者兼而有之。静态结构模型对重大进步做出了贡献，例如在基因编辑技术方面，基于酶‘CRISPR/Cas9’的重新编程。这种结构信息对于药物发现、准确指导设计和优化工作也是至关重要的。这些都是依赖于精确控制蛋白质结构动态变化的主要新应用。通过在时间和空间上连接高分辨率信息，这些目标-以及我们对基础生物学的理解-将大大推进。我的研究将开创一种集成的实验和计算方法，以前所未有的时空分辨率确定酶是如何动态调节的，以及它们如何催化化学反应。我们现在有一个独特的机会来测量大型酶的结构扰动，既有高结构分辨率(每个氨基酸构建块)，也有高时间分辨率(每毫秒)。我们获得的信息将被用来建立高分辨率的动态结构模型，在该模型中，单个特征根据其单独的速率进行重新配置，实验确定的速率为毫秒和氨基酸精度。这项工作将集中在最近在开发生物技术和医学工具方面取得的两个备受瞩目的成功领域，这两个领域依赖于对酶动态结构变化的精确控制。(I)基因编辑：越来越多的人努力改造CRISPR/Cas9酶，以提高它们的效率，并为原位基因组的定向突变创造全新的工具。这在研究中有潜在的广泛应用，但也可以专门用于治疗遗传病。我们将研究基因编辑酶，以提供机制洞察力来解释它们的行为，并指导具有更高活性的变体的开发。(Ii)变构药物的发现。最近，大型制药公司和风险投资/生物技术合作伙伴都进行了重大投资，以发现通过控制蛋白质构象来控制酶功能的“变构”药物。这些药物在选择性和改变疾病中原本难以处理的靶点的能力方面具有潜在的好处。我们将确定我们将开发的新的毫秒时间分辨测量是否可以区分变构调节的信号，从而形成变构药物直接筛选的基础。这项研究计划汇集了建立新的实验方法、尖端数据科学方法、新软件工具开发以及与基础生物学和生物技术和药物发现应用直接相关的专业知识。

The processes of life are dynamic - it is change on a molecular level that enables us to grow and move, but also to become ill and treat disease. Just as the shape and posture of our body can determine our readiness to perform a task, the structure and conformation of a protein molecule can determine its function or activity. It is the ability for proteins to dynamically and rapidly reconfigure that underpins many critical activities in biology, disease and medicine.However, we are currently limited to study proteins, including many important enzymes, at high resolution in space or time - but not both. Static structural models have contributed to major advances, such as in gene editing technology, based on the reprogramming of the enzyme 'CRISPR/Cas9'. This structural information is also crucial for drug discovery, accurately guiding design and optimisation efforts.These are major new applications that rely on precisely controlling dynamic changes in protein structure. These aims - and our understanding of fundamental biology - will be greatly advanced by bridging high resolution information in both time and space.My research will pioneer an integrated experimental and computational approach to determine with unprecedented spatio-temporal resolution how enzymes are dynamically regulated and how they catalyse chemical reactions. We now have a unique opportunity to make measurements of the structural perturbations in large enzymes both with high structural resolution (per amino acid building block) and high temporal resolution (per millisecond). The information that we gain will be used to build high resolution dynamic structural models in which individual features reconfigure according to their individual rates, determined experimentally with millisecond and amino acid precision. This work will focus on two areas of recent high profile success in developing tools for biotechnology and medicine which depend on the exquisite control of enzyme dynamic structural changes. (i) Gene editing: There is a growing effort to engineer CRISPR/Cas9 enzymes to improve their efficiency and to create entirely new tools for targeted mutation of the genome in situ. This has potentially broad application in research, but also to specifically treat genetic diseases. We will study gene editing enzymes to provide mechanistic insight to explain their behaviour and to guide the development of variants with improved activities. (ii) Allosteric drug discovery. There has been major recent investment by both big pharma and by venture capital/biotechnology partnerships to discover 'allosteric' drugs that control enzyme function by controlling the protein conformation. These drugs have potential benefits in selectivity and the ability to modify otherwise intractable targets in disease. We will ascertain whether the new millisecond time-resolved measurements that we will develop can differentiate signatures of allosteric regulation and thus form the basis of a direct screen for allosteric drugs.This research programme brings together expertise in building novel experimental methods, cutting edge data science approaches, development of new software tools and a direct relevance to fundamental biology and applications in biotechnology and drug discovery.