Dynamics, Gating and Opening in Enzyme Catalysis

本申请旨在触及基本但尚未回答的问题的核心，这些问题对于发展我们对酶如何工作的理解至关重要。酶通常是以蛋白质为基础的，它控制着生命系统中几乎所有化学作用的发生速度。了解酶的活性是一个高度优先的问题--它是治疗干预、工业生物技术和合成生物学的核心。酶活性的受控操纵是这些研究领域中的关键因素之一。酶的活性已经被研究了几十年，许多范例已经演变，但最近，已经开发出工具，允许以前所未有的详细程度测试这些范例。在这些工具中，可以说最重要的是核磁共振光谱学，它允许在单个原子的水平上观察酶的结构、电子学和动力学。正确理解所有这些元素及其相互作用对于操纵酶活性至关重要，随着观察力在这一细节水平上的变化，范式正在发生变化。在这项研究中，我们将解决对几乎所有酶的活性都至关重要的问题，即什么控制化学反应的发生速度，是什么让一种酶对另一种反应进行反应，以及是什么决定了它与其作用的化学物质的结合情况。为了实现这一点，我们将研究一种在葡萄糖周围移动磷酸基团的酶的行为。移动磷酸基团的酶位于生物体中每个系统的核心--储存、维护和表达遗传信息、新陈代谢、通讯、细胞结构、分化和动态平衡。我们将根据这种行为开发新的模型，这种原型酶可以提供非常高质量的测量，然后可以转化为治疗、工业生物技术和合成生物学计划中的常见目标酶。

The present application aims to get to the heart of fundamental, yet unanswered, questions that are critical to developing our understanding of how enzymes work. Enzymes, which are normally protein-based, control the rate at which almost all chemistry occurs in living systems. Understanding enzyme activity is a high priority - it is at the core of therapeutic intervention, industrial biotechnology, and synthetic biology. The controlled manipulation of enzyme activity is one of the key elements targeted in each of these areas of research. Enzyme activity has been studied for many decades and many paradigms have evolved but, very recently, tools have been developed that allow the testing of those paradigms with unprecedented levels of detail. Arguably foremost among these tools is NMR spectroscopy, which allows the observation of the structure, electronics and dynamics within enzymes at the level of individual atoms. A proper understanding of all of these elements and their interplay is crucial to manipulating enzyme activity, and with observation powers at this level of detail, paradigms are shifting. In this study we will address questions that are fundamental to the activity of almost all enzymes, namely what controls how fast the chemistry occurs, what makes an enzyme carry out one reaction against another, and what determines how well it binds to the chemicals on which it acts. To achieve this we will study the behaviour of an enzyme that moves phosphate groups around glucose. Enzymes that move phosphate groups lie at the heart of every system in living organisms - in the storage, maintenance and expression of genetic information, in metabolism, communication, cell architecture, differentiation, and homeostasis. We will develop new models based on the behaviour an archetypal enzyme that delivers very high quality measurements, which can then be translated to the enzymes that are common targets in therapeutic, industrial biotechnology and synthetic biology programmes.