Functional Studies of Cytosolic Antioxidant Proteins

描述（由申请人提供）：研究的总体目标 这里提出的是破译氧化还原平衡之间的关系，氧化 真核细胞应激与金属离子代谢及其在人体内的作用 疾病和衰老。特别是，超氧化物应激的详细性质， 也就是说，由于直接化学物质导致氧化应激组分 活细胞内的超氧化物的反应，将寻求。校长 模型系统是芽殖酵母酿酒酵母，但假设 酵母研究中获得的蛋白质也将在小鼠中进行测试。校长 要研究的蛋白质是铜锌超氧化物歧化酶（CuZnSOD）， 一种抗氧化酶及其铜伴侣（CCS），据信 通过插入铜激活CuZnSOD。第一个主要目标是澄清 铁在超氧化物应激中的作用它将通过研究自然 在缺乏CuZnSOD的生物体中积累的游离铁中， 超氧化物使某些含铁蛋白质失活， 遗传抑制因子降低超氧化物敏感性的机制 缺乏CuZnSOD的酵母菌株。第二个主要目标是发现 铜锌代谢、超氧化物胁迫与 CuZnSOD的金属态。研究的目的是了解 CCS法插入CuZnSOD中铜的机理及状态测定 CuZnSOD在不同组织中的代谢率。了解如何使用Metabolic 的CuZnSOD的完成，以及它如何在各种组织中调节将有助于 我们更好地了解人类的抗氧化防御， states.总的来说，这项工作将推进我们对细节的理解。 超氧化物胁迫的化学机制，铁、铜和锌的作用 在超氧化物应激的表现中， 超氧化物应激与人体健康和衰老

DESCRIPTION (provided by applicant): The overall objective of the research proposed here is to decipher the relationships between redox balance, oxidative stress, and metal ion metabolism in eukaryotic cells and their roles in human disease and aging. In particular, the detailed nature of superoxide stress, i.e., that component of oxidative stress which is due to direct chemical reactions of superoxide within living cells, will be sought. The principal model system is the budding yeast Saccharomyces cerevisiae, but hypotheses derived from the yeast studies will be tested in mice as well. The principal proteins to be studied are copper-zinc superoxide dismutase (CuZnSOD), which is an antioxidant enzyme, and its copper chaperone (CCS), which is believed to activate CuZnSOD by inserting copper. The first major goal is to clarify the role of iron in superoxide stress. It will be approached by studying the nature of the free iron that accumulates in organisms lacking CuZnSOD, the inactivation of certain iron-containing proteins by superoxide, and the mechanisms by which genetic suppressors decrease the superoxide sensitivity of yeast strains lacking CuZnSOD. The second major goal is to discover the relationship(s) between copper and zinc metabolism, superoxide stress, and the metallation state of CuZnSOD. Studies will be directed at understanding the mechanism of copper insertion into CuZnSOD by CCS and at determining the state of metallation of CuZnSOD in different tissues. Understanding how metallation of CuZnSOD is accomplished and how it is regulated in various tissues will help us better understand human antioxidant defenses and thus certain disease states. Overall, this work will advance our understanding of the detailed chemical mechanisms of superoxide stress, the roles of iron, copper, and zinc in the manifestation of superoxide stress, and, ultimately, the role of superoxide stress in human health and aging.