THE ROLE OF DYNAMICS IN PEPCK MEDIATED CATALYSIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 糖尿病相关的高血糖是长期糖尿病引起的所有慢性并发症的根本原因。磷酸烯醇式丙酮酸羧激酶(PEPCK)在糖尿病相关高血糖时的血糖升高中起关键作用。我们的研究重点是了解PEPCK如何催化其生物学反应，从而开发出有效的该酶抑制剂作为有效的降血糖药物。我们应用的长期目标是了解蛋白质催化剂固有的动态特性在酶介导的催化过程中所起的作用。这些研究的结果将对设计新的基于蛋白质的催化剂的能力以及提高我们设计有效和选择性的生物重要酶抑制剂治疗疾病的能力具有广泛的深远影响。通过利用磷酸烯醇式丙酮酸羧基激酶，我们的研究将直接导致该酶选择性抑制剂的开发，用于治疗糖尿病相关的高血糖。为了实现这些目标，我们将结合X射线结晶学和核磁共振的优势，不仅表征催化过程中发生的构象变化，而且还表征这些运动发生的时间尺度。通过将这些结构和动力学研究与酶在野生型酶和被设计来颠覆动态过程的突变体中进行催化的能力相关联，我们将更深入地了解这些动态变化在催化循环中所起的特殊作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Diabetes-associated hyperglycemia is the underlying cause of all of the chronic complications arising from a prolonged diabetic condition. The enzyme phosphoenolpyruvate carboxykinase (PEPCK) plays a key role in the elevated blood glucose levels observed during diabetes-associated hyperglycemia. Our studies focus on understanding how PEPCK catalyzes its biological reaction will lead to the development of effective inhibitors of this enzyme as potent anti-hyperglycemic agents. Our application's long-term goals are to understand the role that the inherent dynamic properties of protein catalysts play in the process of enzyme mediated catalysis. The results of these studies will have broad reaching implications in the ability to design novel protein based catalysts as well as advance our ability to design potent and selective inhibitors of biologically important enzymes in the treatment of disease. By utilizing the enzyme phosphoenolpyruvate carboxykinase our studies will lead directly to the development of selective inhibitors of this enzyme in the treatment of diabetes-associated hyperglycemia. To achieve these goals, we will combine the strengths of x-ray crystallography and NMR to characterize not only the conformational changes that occur during catalysis, but also the time scale in which these motions occur. By correlating these structural and dynamics studies with the ability of the enzyme to carry out catalysis in both the wild-type enzyme and in mutants engineered to upset the dynamic processes, we will achieve a more thorough understanding of the particular roles that these dynamic changes play in the catalytic cycle.