A Biomimetic Approach to Elucidating the Role in Disease of the Oxygenase Enzyme Acireductone Dioxygenase (ARD)

阐明加氧酶 Acireductone 双加氧酶 (ARD) 在疾病中的作用的仿生方法

• 批准号: 10392383
• 负责人: Santiago Andres Toledo Carrion
• 金额: $ 9.54万
• 依托单位: ST. EDWARD'S UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392383
• 关键词: Active Sites; Address; Animals; Apoptotic; Bacteria; Binding; Biological Models; Biomimetics; Brain Neoplasms; Carbon Monoxide; Catalysis; Cell Proliferation; Cells; Chemistry; Cobalt; Code; Complex; Copper; DNA biosynthesis; Dependence; Development; Dioxygenases; Disease; Environment; Enzymes; Ethylenes; Family; Genes; Genetic study; Growth; Homologous Gene; Human; In Vitro; Inorganic Chemistry; Investigation; Ions; Iron; Kinetics; Knowledge; Laboratories; Ligands; Manganese; Metals; Methionine; Mission; Modeling; Mus; Nickel; Oncogenic; Oxygen; Oxygenases; Pathology; Pathway interactions; Plants; Play; Polyamines; Propionates; Prostate; Rattus; Reaction; Regulation; Role; S-Adenosylhomocysteine; Signaling Molecule; Structure; System; Testing; United States National Institutes of Health; Work; analog; base; cancer cell; cell growth; design; fibrosarcoma; in vivo; inhibitor; metalloenzyme; non-Native; oxidation; prevent; prostate cancer cell; small molecule; transmethylation; tumor

Project Summary/Abstract Acireductone dioxygenase (ARD) is found in the ubiquitous methionine salvage pathway (MSP) in animals, plants, and bacteria. Enzymatically it is implicated in the regulation of SAM and MTA; the latter is a result of polyamine synthesis and a regulator of cell growth in animals. The enzyme can bind to iron and nickel giving two distinct products. The nickel reaction (“off-pathway”) catalyzes the transformation of acireductone into 3- (methylthio)propionate, formate and carbon monoxide, a known anti-apoptotic signaling molecule. ARD is the only known example of a metalloenzyme whose function differs only by the identity of the metal ion. Recent mammalian studies have characterized mouse (MmARD) and human analogues (HsARD) of the enzyme. In mammalian systems it has been shown that ARD is capable of binding manganese and cobalt to perform “off- pathway” type chemistry. Recent work with these analogues in in-vitro studies has shown that HsARD might play an intracellular regulatory role in brain tumors. Furthermore, it has been shown that the gene coding for HsARD, ADI1, is downregulated in human and rat prostate cancer cells as well as gastrocarcinoma and fibrosarcoma cells. These moonlighting functions, as well as the enzymatic roles of ARD beg the question about the connection between ARD in disease. To date there is still debate about the exact mechanism of the regiospecific substrate oxidation, and little is known about the role that metal identity plays in this reactivity. This project uses synthetic biomimetic modeling to contribute to answering these questions. The proposed work aims at greatly expanding the availability of models of ARD using nickel and mammalian relevant metals with two specific aims: 1. Expand the limited availability of biomimetic structural and functional models of Ni- ARD. These will be used to test the structural and mechanistic reasons for the observed “off-pathway” oxidative regioselective reactivity in ARD. Mechanistic and kinetic studies will follow to provide valuable information to explain the mechanism of regioselective substrate activation in Ni-ARD. 2. Synthesize and begin to study the biomimetic reactivity of model complexes using metals relevant to mammalian systems (cobalt and manganese specifically). These new compounds will contribute to the understanding of the “off-pathway” reactions and the role of metal identity in promoting disease in human cells.

项目总结/摘要 酸还原酮双加氧酶（ARD）存在于动物体内普遍存在的蛋氨酸补救途径（MSP）中， 植物和细菌。在酶促作用下，它参与SAM和MTA的调节;后者是 多胺合成和动物细胞生长的调节剂。这种酶可以与铁和镍结合， 两种不同的产品。镍反应（“非途径”）催化酸还原酮转化为3-羟基-3-甲基-苯并[d]吡啶。 （甲硫基）丙酸酯、甲酸酯和一氧化碳，一种已知的抗凋亡信号分子。ARD是 唯一已知的金属酶的例子，其功能的不同之处仅在于金属离子的身份。最近 哺乳动物研究已经表征了该酶的小鼠（MmARD）和人类似物（HsARD）。在 在哺乳动物系统中，已经显示ARD能够结合锰和钴以执行“关闭”， “途径”型化学。最近在体外研究中对这些类似物的研究表明，HsARD可能 在脑肿瘤中发挥细胞内调节作用。此外，已经表明，编码 HsARD，ADI 1，在人类和大鼠前列腺癌细胞以及胃癌中下调， 纤维肉瘤细胞这些兼职的功能，以及ARD的酶的作用提出了一个问题， ARD与疾病之间的联系。到目前为止，关于这种现象的确切机制仍有争议。 区域特异性的基板氧化，并很少有人知道的作用，金属身份在这种反应性。 这个项目使用合成仿生建模来回答这些问题。拟议 这项工作的目的是利用镍和哺乳动物相关的金属，极大地扩展ARD模型的可用性 有两个具体目标：1。扩大镍的仿生结构和功能模型的有限可用性- ARD。这些将被用来测试所观察到的“偏离路径”的结构和机制原因 ARD中的氧化区域选择性反应性。机制和动力学研究将遵循提供有价值的 信息来解释在Ni-ARD中区域选择性底物活化的机制。2.合成并开始 为了研究模型络合物的仿生反应性，使用与哺乳动物系统相关的金属（钴和 锰）。这些新化合物将有助于理解“关闭途径” 反应和金属身份在促进人类细胞疾病中的作用。