PHYSICAL/CHEMICAL TRAPPING & LAUE DIFFRACTION/TIME-RESOLVED CRYSTALLOGRAPHY O

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。 摘要：酶在催化的不同阶段的可视化对于理解酶的催化机制是非常有价值的，并已被成功地用作合理的药物设计的工具。模拟底物及其在催化过程中的结构变化是设计抑制剂和药物的有效手段。事实上，大量的药物都是目标酶--S底物、产物、中间体或过渡态的模拟类似物。对产生的底物复合体的直接观察也提供了关于蛋白质动力学的有价值的信息。酶反应是酶及其配体结构变化的流体连续体。在酶学、蛋白质工程和合理的药物设计方面的最新发展需要动态的结构信息来理解酶反应的整个连续体，并拓宽药物的潜在靶点。我们建议使用三种疾病相关的酶模型，弹性酶，N-酰基高丝氨酸内酯水解酶和天冬氨酸氨基转移酶来探索酶催化的结构动力学，以期引导新的药物设计。我们计划使用不同的化学/物理捕获方法来捕捉酶活性部位的瞬时结构性质。此外，我们希望尽最大努力开发一种时间分辨系统来研究一般的酶反应。

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. Abstract: The visualization of enzymes trapped in various stages of catalysis has been invaluable in understanding the catalytic mechanisms of enzymes and has been used successfully as a tool for rational drug design. Mimicry of a substrate and its transformed structures during catalysis is an effective means to design inhibitors and drugs. Indeed, a great number of drugs are mimetic analogs of the target enzyme?s substrates, products, intermediates, or transition state. Direct observation of the productive substrate complexes has also provided valuable information about protein dynamics. An enzymatic reaction is a fluid continuum of structural changes of both the enzyme and its ligands. Recent developments in enzymology, protein engineering, and rational drug design demand dynamic structural information to understand the whole continuum of an enzymatic reaction and to broaden the potential targets of drugs. We propose to use three diseases-related enzyme models, elastase, N-acyl homoserine lactone hydrolas, and aspartate aminotransferase to explore the structural dynamics of enzyme catalysis in hope of leading to novel drug design. We plan to use different chemical/physical trapping methods to capture the transient structural natures of enzyme active sites. Furthermore, we would like to make our best effort to develop a time-resolved system to study enzymatic reactions in general.