Copper Transport Mechanism of Menkes disease protein and Wilson disease protein

描述（由申请人提供）：铜是人体正常代谢所必需的。体内分布铜的蛋白质功能的先天性缺陷导致严重的代谢紊乱，例如门克斯病和威尔逊病，其特征在于神经和肝脏异常、肾和血管损伤。铜转运ATP酶，ATP 7A（门克斯病蛋白）和ATP 7 B（威尔逊病蛋白），已被确定为人类细胞中铜浓度的关键调节因子。近年来，这些转运蛋白的生物化学和细胞内特性的信息已经迅速产生。然而，关于ATP 7A和ATP 7 B的不同生理作用知之甚少。我们的长期目标是表征ATP 7A和ATP 7 B在铜稳态中的特定作用，作为开发和改善这些疾病的治疗方法的先决条件。本提案的主要目标是了解ATP 7A和ATP 7 B在铜转运中的具体作用。使用重组蛋白的建设，定点诱变，片段交换实验，铜结合亲和力测定，我们将确定一个独特的序列插入的作用，在功能和交通的ATP 7A和ATP 7 B。此外，我们将研究受体蛋白对铜释放的作用，通过测量过表达或敲低肽基-a-单加氧酶后ATP 7A的铜转运速率。最后，我们将研究 ATP 7 B活性对CIC-4的依赖性，CIC-4是一种哺乳动物内体Cl-/H+交换剂，在细胞生理学中的功能知之甚少。这项研究的结果将为了解和更好地治疗门克斯病和威尔逊病提供必要的信息。 公共卫生相关性：Menkes病和Wilson病的特征在于神经和肝脏异常、肾和血管损伤，并且分别由酶ATP 7A和ATP 7 B的功能破坏引起。我们的目标是表征ATP 7A和ATP 7 B的特定作用，作为开发和改善这些疾病治疗方法的先决条件。

DESCRIPTION (provided by applicant): Copper is essential for normal human metabolism. Inborn defects in function of proteins distributing copper within the body lead to severe metabolic disorders, such as Menkes disease and Wilson disease, which are characterized by neurological and hepatic abnormalities, renal and vascular damage. The copper transporting ATPases, ATP7A (Menkes disease protein) and ATP7B (Wilson disease protein), have been identified as key regulators of copper concentration in human cells. In recent years, information on biochemical and intracellular properties of these transporters has been rapidly generated. However, little is known about the distinct physiological roles of ATP7A and ATP7B. Our long term goal is to characterize specific roles of ATP7A and ATP7B in copper homeostasis as a prerequisite to the development and improvement of therapeutic treatments for these disorders. The major goal of this proposal is to understand specific roles of ATP7A and ATP7B in copper transport. Using recombinant protein construction, site directed mutagenesis, segment exchanging experiments, and copper binding affinity assays; we will determine the role of a unique sequence insert in the function and trafficking of ATP7A and ATP7B. Also, we will investigate the role of acceptor proteins on copper release by measuring the rate of copper transport of ATP7A after overexpression or knock-down of peptidyl-a-monooxygenase. Finally, we will examine the dependence of ATP7B activity on CIC-4, a mammalian endosomal CI-/H+ exchanger with poorly understood function in cell physiology. The result of the proposed research will yield information essential for understanding and better treatment of Menkes disease and Wilson disease. PUBLIC HEALTH RELEVANCE: Menkes disease and Wilson disease are characterized by neurological and hepatic abnormalities, renal and vascular damage, and are caused by a disruption in function of the enzymes ATP7A and ATP7B, respectively. Our goal is to characterize the specific roles of ATP7A and ATP7B as a prerequisite to the development and improvement of therapeutic treatments for these disorders.