Using Structural Studies to Investigate the Relationships Between Enzymes and Ligands.

利用结构研究研究酶和配体之间的关系。

• 批准号: RGPIN-2022-05150
• 负责人: Sanders, David
• 金额: $ 2.48万
• 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: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748078
• 关键词: Using; Structural; Studies; Investigate; Relationships

My research is focused on using protein structures to understand how molecules recognize each other in biological systems. My lab has established its reputation by using structural studies to enhance understanding of the interactions between enzymes and their substrates. This allows us to predict the effects that changes in the interactions will have on enzymes. Combining crystallography with enzymology, computational studies, and other biophysical techniques, we can characterize the ways that ligands interact with proteins. We have the long-term goal of using this information to develop compounds that can affect enzyme processes and to understand the evolution of enzyme systems. We are using the thioredoxin system as a model system for studying protein-protein interactions. The thioredoxin system is a critical system in oxidative stress. Our studies have increased our understanding of the interactions between thioredoxin (Trx) and thioredoxin reductase (TrxR). We predict that we can design peptides that exhibit specificity towards TrxR or Trx. Structural studies, enzymology, and other techniques to monitor protein-peptide binding (eg ITC and SPR) will help improve peptide binding. Understanding the evolution of enzyme structure-function relationships remains a key focus of my lab. We will continue to use the thioredoxin system to continue to study the way Trx and TrxR interact under extreme conditions. We have chosen this system because the chemistry is very well characterized, the Trx system is ubiquitous and oxidative stress is of great scientific interest. Using crystallography and enzymology we will further characterize the interactions of the thioredoxin system of extremophiles. 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 recognized by enzymes and how this binding affects enzyme behavior. Dihydrodipicolinate synthase(DHDPS) is regulated by lysine at an allosteric site. We have determined structures of DHDPS in the presence and absence of substrates and inhibitors, to study how the allosteric inhibition of this enzyme occurs. Our hypothesis is that inhibitor-induced changes in flexibility are responsible for the loss in activity. We will use bioinformatics, enzymology, and molecular dynamic studies to target specific amino acids to probe the flexible/rigid regions in controlling allosteric inhibition. These studies will greatly increase our knowledge in how flexibility changes affect the behavior of enzymes. Our access to state-of-the-art facilities, including the CVS, the SSSC, and the PCCF located at the University of Saskatchewan enables us to be leaders in studying protein structure-function relationships and over the next five years we will continue to investigate novel systems to address important questions about protein:ligand interactions.

我的研究重点是利用蛋白质结构来了解分子如何在生物系统中相互识别。我的实验室通过使用结构研究来增强对酶与其底物之间相互作用的理解而建立了声誉。这使我们能够预测相互作用的变化对酶的影响。结合晶体学与酶学，计算研究和其他生物物理技术，我们可以表征配体与蛋白质相互作用的方式。我们的长期目标是利用这些信息开发可以影响酶过程的化合物，并了解酶系统的进化。 我们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。硫氧还蛋白系统是氧化应激的关键系统。我们的研究增加了我们对硫氧还蛋白（Trx）和硫氧还蛋白还原酶（TrxR）之间相互作用的理解。我们预测，我们可以设计对TrxR或Trx表现出特异性的肽。结构研究、酶学和其他监测蛋白质-肽结合的技术（如ITC和SPR）将有助于改善肽结合。 了解酶结构-功能关系的演变仍然是我实验室的一个重点。我们将继续使用硫氧还蛋白系统，继续研究极端条件下Trx和TrxR相互作用的方式。我们之所以选择这个系统，是因为它的化学特性非常好，Trx系统无处不在，氧化应激具有很大的科学意义。利用晶体学和酶学，我们将进一步表征极端微生物硫氧还蛋白系统的相互作用。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酶如何识别小分子以及这种结合如何影响酶的行为。二氢吡啶二羧酸合酶（DHDPS）在变构位点受赖氨酸调节。我们已经确定了在存在和不存在的底物和抑制剂的DHDPS的结构，研究如何发生这种酶的变构抑制。我们的假设是，走廊引起的灵活性的变化是负责活动的损失。我们将使用生物信息学，酶学和分子动力学研究，以特定的氨基酸为目标，探测控制变构抑制的柔性/刚性区域。这些研究将大大增加我们对灵活性变化如何影响酶行为的认识。我们获得了最先进的设施，包括CVS，SSSC和位于萨斯喀彻温大学的PCCF，使我们能够成为研究蛋白质结构-功能关系的领导者，在未来五年内，我们将继续研究新的系统，以解决有关蛋白质的重要问题：配体相互作用。