Understanding enzyme-substrate recognition in catalysis and inhibition.

了解催化和抑制中的酶-底物识别。

• 批准号: RGPIN-2016-05765
• 负责人: Sanders, David
• 金额: $ 2.4万
• 依托单位: University of Saskatchewan
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=615891
• 关键词: Understanding; enzyme; substrate; recognition; catalysis

Research in my lab is focused on the broad theme of using protein X-ray crystallography to understand molecular recognition in enzyme systems. By combining crystallography with enzymology and other biophysical techniques, we can characterize the roles of individual amino acids in ligand recognition. We are using state of the art facilities, including the Canadian Light Source, the Saskatchewan Structural Sciences Centre, and the Protein Characterization and Crystallization Facility located at the University of Saskatchewan. My lab has established its reputation by using structural studies to enhance our understanding of how enzymes recognize and bind their ligands. By examining these interactions at the atomic level, we can predict the effects of alterations on binding. We have the ultimate goal of using the information to develop compounds that can more effectively interact with enzymes and to use structural information to predict substrates for new, poorly understood enzymes. We have been using the thioredoxin system as a model system for studying protein-protein interactions. Our current research objectives are to probe the interactions between thioredoxin (Trx) and thioredoxin reductase (TrxR), to characterize the interactions that are important for binding and to examine the way these enzymes are compensating for temperature differences and other evolutionary pressures to maintain their interaction. We have chosen this system because the chemistry is very well characterized, the Trx system is ubiquitous and oxidative stress (in which Trx plays a regulatory role) is of great current scientific interest. By using crystallography and enzymology we are characterizing the interactions of the thioredoxin system from extremophiles to determine the recognition characteristics of this system. A full understanding of the interactions that determine the recognition and binding of the TrxR/Trx complex will offer insights into other protein-protein interactions. Our work also involves understanding how small molecules are specifically recognized by enzymes. In past grants, we have studied protein-ligand interactions for a variety enzymes, including inositol dehydrogenase (IDH), dihydrodipicolinate synthase (DHDPS) and UDP-galactopyranose mutase (UGM). We have determined the structures of these enzymes complexed with substrates and inhibitors in order to determine the modes of interactions and substrate recognition. We have utilized this information for to give us insights into how these diverse enzymes catalyze their reactions. Additionally, the information gained from our structural studies have aided us in identifying new sub-classes of enzyme families and to design novel enzyme inhibitors. In this grant, we propose to continue our work in using protein x-ray crystallography and enzymology to examine the interactions between novel enzymes and their ligands.

我实验室的研究集中在使用蛋白质X射线晶体学来理解酶系统中的分子识别的广泛主题上。通过结合晶体学与酶学和其他生物物理技术，我们可以表征单个氨基酸在配体识别中的作用。我们正在使用最先进的设施，包括加拿大光源，萨斯喀彻温省结构科学中心，以及位于萨斯喀彻温大学的蛋白质表征和结晶设施。 我的实验室已经建立了它的声誉，通过使用结构研究，以提高我们对酶如何识别和结合其配体的理解。通过在原子水平上检查这些相互作用，我们可以预测改变对结合的影响。我们的最终目标是利用这些信息来开发能够更有效地与酶相互作用的化合物，并利用结构信息来预测新的、知之甚少的酶的底物。 我们一直使用硫氧还蛋白系统作为研究蛋白质-蛋白质相互作用的模型系统。我们目前的研究目标是探索硫氧还蛋白（Trx）和硫氧还蛋白还原酶（TrxR）之间的相互作用，表征的相互作用是重要的结合，并检查这些酶是如何补偿温度差异和其他进化压力，以保持其相互作用。我们之所以选择这个系统，是因为其化学特性非常好，Trx系统无处不在，氧化应激（其中Trx起着调节作用）是当前最受关注的科学。通过使用晶体学和酶学，我们正在表征的硫氧还蛋白系统从极端微生物的相互作用，以确定该系统的识别特性。对决定TrxR/Trx复合物的识别和结合的相互作用的充分理解将提供对其他蛋白质-蛋白质相互作用的见解。 我们的工作还包括了解小分子如何被酶特异性识别。在过去的赠款中，我们研究了各种酶的蛋白质-配体相互作用，包括肌醇脱氢酶（IDH），二氢吡啶二羧酸合酶（DHDPS）和UDP-吡喃半乳糖苷酶（UGM）。我们已经确定了这些酶与底物和抑制剂复合的结构，以确定相互作用和底物识别的模式。我们已经利用这些信息来让我们深入了解这些不同的酶如何催化它们的反应。此外，从我们的结构研究中获得的信息有助于我们识别酶家族的新子类和设计新的酶抑制剂。在这项资助中，我们建议继续我们的工作，利用蛋白质x射线晶体学和酶学研究新的酶和它们的配体之间的相互作用。