ENZYMES OF THE PROSTAGLANDIN CASCADE

前列腺素级联酶

• 批准号: 6109702
• 负责人: R Michael Garavito
• 金额: --
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6109702
• 关键词: X ray crystallography; chemical structure function; circular dichroism; eicosanoid metabolism; electron density; enzyme biosynthesis; enzyme structure; isomerase; ligands; membrane proteins; molecular pathology; prostaglandin endoperoxide synthase; prostaglandins; protein purification; protein structure function; recombinant proteins; sickle cell anemia; vasoconstrictors

The pathological manifestations of sickle cell anemia arises from the polymerization of HbS into long fibers which distort the shape of the red blood cell. The resulting abnormal shape and increased rigidity of the red blood cells impedes blood flow through the capillaries causing the vasoocclusive complications that characterize sickle cell anemia. Cerebral vascular disease is a common and one of many catastrophic complications in children suffering from sickle cell anemia. We have started a long-term investigation into the structure and functional of enzymes in the prostaglandin synthesis cascade. Most of these enzymes are membrane bound enzymes and thus require special handing to prepare them for biophysical analysis. As we have developed methods for crystallizing membrane proteins, they now can be made amendable to X-ray diffraction analysis. Using X- ray crystallography, electron paramagnetic resonance, UV-Vis absorption spectroscopy and fluorine-NMR, we intend to study the enzymes that participate in the synthesis of vasoregulatory prostaglandins. We are now investigating enzyme-drug and enzyme- substrate analog complexes of prostagland H synthase (cyclo- oxygenase), both in solution and in crystals. The products of prostaglandin H synthase, PGH1 and PGH2, serve as precursors for synthesis of prostaglandins which affect platelet aggression and vasodilation. We intend to use the methods we have developed for the study prostaglandin synthase to purify, crystalize and investigate and investigate PGH-PGE isomerase and PGI2 synthase which synthesize powerful vasodilators and inhibitors of platelet aggregation. Our long term goal is to elucidate the structural mechanisms of substrate, product and inhibitor binding to 1) understand better the molecular events occurring during catalysis and 2) to provide the necessary structural information for the design of drugs to control the synthesis of specific prostaglandin. Having inhibitors or activators of the synthesis of specific prostaglandins could allow the development of rational drug therapy for vasoocclusive diseases like sickle cell anemia.

镰状细胞性贫血的病理表现源于 HbS聚合成长纤维， 红细胞的数量 由此产生的异常形状和增加 红细胞的刚性阻碍血液流过 毛细血管引起血管闭塞并发症， 镰状细胞性贫血的特征 脑血管病是一种 儿童常见的灾难性并发症之一 患有镰状细胞性贫血 我们已经开始对该结构进行长期调查， 前列腺素合成级联中酶的功能。 最 这些酶中的一种是膜结合酶，因此需要 特殊处理，为生物物理分析做好准备。 正如我们 已经开发了使膜蛋白结晶的方法， 现在可以进行X射线衍射分析。 使用X- 射线晶体学，电子顺磁共振，紫外-可见 吸收光谱和氟核磁共振，我们打算研究 参与血管调节物质合成的酶 兰丁 我们现在正在研究酶-药物和酶- 底物类似物复合物的H2O合酶（环， 加氧酶），在溶液和晶体中。 的产品 前列腺素H合酶，PGH 1和PGH 2，作为前列腺素合成酶的前体， 影响血小板攻击性和 血管舒张 我们打算使用我们开发的方法， 前列腺素合成酶纯化、结晶及 PGH-PGE异构酶和PGI 2合酶研究 合成强效血管扩张剂和血小板抑制剂 聚合来 我们的长期目标是阐明 底物、产物和抑制剂结合的机制1） 更好地理解催化过程中发生的分子事件 和2）提供必要的结构信息， 设计控制特定前列腺素合成的药物。 具有特异性合成的抑制剂或激活剂的 三尖杉酯碱可以促进合理药物治疗的发展 治疗血管闭塞性疾病如镰状细胞性贫血