Copper Transport Mechanism of Menkes disease protein and Wilson disease protein

门克斯病蛋白和威尔逊病蛋白的铜转运机制

• 批准号: 7707973
• 负责人: Amanda Barry
• 金额: $ 4.72万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-29 至 2011-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7707973
• 关键词: ATP phosphohydrolase; ATP7A protein; Affinity; Anabolism; Apical; Behavior; Binding; Biochemical; Biological Assay; Brain; Calorimetry; Catalysis; Cell membrane; Cell physiology; Cells; Ceruloplasmin; Characteristics; Chloride Ion; Chlorides; Copper; Cytosol; Data; Defect; Dependence; Development; Disease; Down-Regulation; Endocytic Vesicle; Engineering; Enzymes; Goals; Golgi Apparatus; Growth and Development function; Hepatic; Hepatocyte; Hepatolenticular Degeneration; Homeostasis; Human; In Vitro; Insecta; Laboratories; Lead; Life; Measures; Membrane; Menkes Kinky Hair Syndrome; Metabolic; Metabolic Diseases; Metabolism; Mixed Function Oxygenases; Neurologic; Organism; Pathway interactions; Phosphorylation; Physiological; Placenta; Property; Protein Dephosphorylation; Proteins; Recombinant Proteins; Regulation; Research; Research Training; Role; Scaffolding Protein; Site-Directed Mutagenesis; Testing; Therapeutic; Titrations; Vesicle; Wilson disease protein; base; basolateral membrane; cell type; cofactor; copper-transporting ATPase; extracellular; kidney vascular structure; knock-down; meetings; mutant; overexpression; polarized cell; protein function; public health relevance; research study; trafficking

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.

描述（由申请人提供）：铜对于人体正常新陈代谢至关重要。体内分配铜的蛋白质功能的先天缺陷会导致严重的代谢紊乱，例如门克斯病和威尔逊病，其特征是神经和肝脏异常、肾脏和血管损伤。铜转运 ATP 酶 ATP7A（门克斯病蛋白）和 ATP7B（威尔逊病蛋白）已被确定为人体细胞中铜浓度的关键调节因子。近年来，有关这些转运蛋白的生化和细胞内特性的信息已迅速产生。然而，人们对 ATP7A 和 ATP7B 的不同生理作用知之甚少。我们的长期目标是确定 ATP7A 和 ATP7B 在铜稳态中的具体作用，作为开发和改进这些疾病的治疗方法的先决条件。该提案的主要目标是了解 ATP7A 和 ATP7B 在铜运输中的具体作用。使用重组蛋白构建、定点诱变、片段交换实验和铜结合亲和力测定；我们将确定独特的序列插入在 ATP7A 和 ATP7B 的功能和运输中的作用。此外，我们将通过测量肽基-α-单加氧酶过度表达或敲低后 ATP7A 的铜转运速率来研究受体蛋白对铜释放的作用。最后，我们将检查 ATP7B 活性对 CIC-4 的依赖性，CIC-4 是一种哺乳动物内体 CI-/H+ 交换器，其在细胞生理学中的功能知之甚少。拟议研究的结果将为理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：门克斯病和威尔逊病的特点是神经和肝脏异常、肾脏和血管损伤，分别是由 ATP7A 和 ATP7B 酶功能破坏引起的。我们的目标是描述 ATP7A 和 ATP7B 的具体作用，作为开发和改进这些疾病治疗方法的先决条件。