Tethered Domains As Regulatory Elements

描述(申请人提供)：我们的工作有助于发现ACT结构域，这是一个由小分子调节的蛋白质调节域，并产生了典型的ACT结构域的结构。这项研究的目的是继续我们对ACT结构域对蛋白质调控机制的研究，并将其与本实验室的长期目标联系起来，该实验室的长期目标是确定构象调控机制中结构和功能之间的关系，最终将其与这些酶和类似系统介导的生理学和病理生理学联系起来，并最终充分了解蛋白质中的构象中介过程，以便能够将基本原理应用于更复杂的系统。这些特异的目的将1)阐明大肠杆菌和结核分枝杆菌D-3-磷酸甘油脱氢酶(PgDn)中ACT结构域的作用机制，2)阐明在结核分枝杆菌中发现的一个新的结构域和小分子结合位点的功能，它可能与ACT结构域一起发挥作用，3)确定人PGDH的结构和功能，以及4)筛选结核分枝杆菌PGDH的配体结合部位，以开发可用于评估PGDH在结核分枝杆菌活性代谢过程中的作用的抑制剂。适应期和持续期，这种药物最终可能用于治疗耐多药结核病。这项工作的相关性是2倍的。首先，ACT结构域是一个新发现的结构域，在许多蛋白质的调节中发挥作用。确定其机制及其在蛋白质中作用的普遍性，将有助于我们理解蛋白质在生理和疾病中的调控，并为基因替代治疗中的蛋白质调控提供一种潜在的新武器。其次，耐多药结核病是一个日益严重的问题，影响到世界上很大一部分人口。我们观察到结核分枝杆菌的前列腺素脱氢酶受L-丝氨酸的抑制，而人的前列腺素脱氢酶不受抑制，这可能为治疗结核病的药物开发提供一个新的焦点。

DESCRIPTION (provided by applicant): Our work has contributed to the discovery of the ACT domain, a protein regulatory domain modulated by small molecules, and has produced the structure of the archetypical ACT domain. The goal of this research is to continue our investigation into the mechanism of regulation of proteins by the ACT domain and to relate this to the long term goals of this laboratory which are to determine the relationship between structure and function in conformationally regulated control mechanisms, to eventually relate that to the physiology and pathophysiology mediated by these enzymes and by analogous systems, and to eventually understand conformational mediated processes in proteins well enough to be able to apply basic principles to more complex systems. The specific aims will 1) elucidate the mechanism of the ACT domain in D-3- phosphoglycerate dehydrogenase (PGDN) from E. coli and M. tuberculosis, 2) elucidate the function of a new structural domain and small molecule binding site found in M. tuberculosis PGDH that may function in conjunction with the ACT domain, 3) determine the structure and function of human PGDH, and 4) screen the ligand binding sites of M. tuberculosis PGDH for the development of inhibitors that can be used to assess the role of PGDH in the metabolism of M. tuberculosis during its active, adaptive and persistent phases and that may eventually be used for the treatment of multi-drug resistant tuberculosis. The relevance of this work is 2-fold. First, the ACT domain is a newly recognized structural domain that functions in the regulation of many proteins. Determining its mechanism and the universality of its action among proteins will contribute to our understanding of protein regulation in physiology and disease as well as provide a potential new weapon for protein regulation in gene replacement therapy. Secondly, multi-drug resistant tuberculosis is a growing problem that impacts a large percentage of the world's population. Our observations that PGDH from M. tuberculosis is inhibited by L-serine and that PGDH from humans is not, may provide a new focus in drug development for treatment of tuberculosis.