STRUCTURE OF THE NUCLEOTIDE-BINDING DOMAINS OF HUMAN COPPER-TRANSPORTING ATPASES

人铜转运ATP酶的核苷酸结合域的结构

• 批准号: 7954614
• 负责人: OLEG Y DMITRIEV
• 金额: $ 0.02万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954614
• 关键词: ATP phosphohydrolase; ATP7A protein; Binding; Binding Sites; Biochemical; Ca(2+)-Transporting ATPase; Computer Retrieval of Information on Scientific Projects Database; Copper; Data Collection; Disease; Environment; Enzymes; Funding; Future; Genes; Goals; Grant; Heavy Metals; Hepatolenticular Degeneration; Human; Inborn Errors of Metabolism; Institution; Location; Metabolism; Mutation; N Domain; Nucleotides; Process; Property; Proteins; Publishing; Regulation; Research; Research Personnel; Resolution; Resources; Solutions; Source; Structure; Surface; Transmembrane Transport; United States National Institutes of Health; Variant; Walkers; Wilson disease protein; copper-transporting ATPase; disease-causing mutation; domain mapping; human tissue; mutant; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of this project is to solve the structures of the nucleotide binding domains of the two human copper-transporting ATPases, Wilson disease associated protein and Menkes disease associated protein and to elucidate conformational changes caused in the domains by ATP binding. The copper-transporting ATPases belong to a distinct and poorly studied class of membrane transport enzymes (Lutsenko et al., 2002) and are essential for copper-metabolism in human tissues. Mutations in the ATP7A and ATP7B genes encoding these proteins result in severe hereditary metabolic disorders. No high-resolution structure of a heavy metal transporting ATPase has been published to date.The structures of the nucleotide binding domains will be solved in the nucleotide-free and ATP-bound forms. Location of the ATP -binding site in the three-dimensional structure of the domains will be determined. Conformational changes taking place upon ATP binding will be studied in the context of the catalytic cycle of the P-type ATPases, with the emphasis on unique features separating heavy-metal transporting ATPases from well studied Ca2+-ATPase. The differences in the three-dimensional structures of the nucleotide binding domains of Wilson disease and Menkes disease proteins associated with non-homologous regions of primary structure will be determined. This project will also provide groundwork for the future studies of the structural effects of disease-causing mutations in the copper-transporting ATPases, for solving the structures of the larger domains of Wilson and Menkes disease proteins and mapping the domain-domain interaction surfaces. To date the structure of the N-nucleotide-binding domain of the Wilson disease ATPase (WNABD) comprising residues 1036-1196 has been solved in ATP bound form (Dmitriev et al., 2006). Data collection and processing for structure determination of two disease-causing mutant variants of the N-domain (E1064A and H1069A) is in progressReferences 1. Dmitriev, O., Tsivkovskii, R., Abildgaard, F., Morgan, C. T., Markley, J. L., and Lutsenko, S. (2006) Solution structure of the N-domain of Wilson disease protein: Distinct nucleotide-binding environment and effects of disease mutations Proc. Natl. Acad. Sci. U. S. A 103, 5302-5307. 2. Lutsenko, S., Efremov, R. G., Tsivkovskii, R., and Walker, J. M. (2002) Human copper-transporting ATPase ATP7B (the Wilson's disease protein): biochemical properties and regulation J. Bioenerg. Biomembr. 34, 351-362.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 该项目的目标是解析两种人类铜转运 ATP 酶、威尔逊病相关蛋白和门克斯病相关蛋白的核苷酸结合域的结构，并阐明 ATP 结合在这些域中引起的构象变化。铜转运 ATP 酶属于一类独特且研究较少的膜转运酶 (Lutsenko et al., 2002)，对于人体组织中的铜代谢至关重要。编码这些蛋白质的 ATP7A 和 ATP7B 基因突变会导致严重的遗传性代谢紊乱。迄今为止，尚未公布重金属转运 ATP 酶的高分辨率结构。核苷酸结合域的结构将以无核苷酸和 ATP 结合的形式得到解决。 将确定 ATP 结合位点在域的三维结构中的位置。 将在 P 型 ATP 酶催化循环的背景下研究 ATP 结合时发生的构象变化，重点是区分重金属转运 ATP 酶与经过充分研究的 Ca2+-ATP 酶的独特特征。将确定与一级结构的非同源区域相关的威尔逊病和门克斯病蛋白的核苷酸结合结构域的三维结构的差异。该项目还将为未来研究铜转运 ATP 酶中致病突变的结构效应、解决威尔逊和门克斯疾病蛋白较大结构域的结构以及绘制结构域-结构域相互作用表面提供基础。 迄今为止，包含残基 1036-1196 的威尔逊病 ATP 酶 (WNABD) 的 N-核苷酸结合结构域的结构已以 ATP 结合形式得到解析 (Dmitriev 等，2006)。正在收集和处理 N 结构域的两种致病突变变体（E1064A 和 H1069A）的结构确定参考文献 1. Dmitriev, O.、Tsivkovskii, R.、Abildgaard, F.、Morgan, C. T.、Markley, J. L. 和 Lutsenko, S. (2006) 威尔逊病蛋白 N 结构域的溶液结构：不同核苷酸结合环境和疾病突变的影响Proc。国家。阿卡德。科学。美国 A 103, 5302-5307。 2. Lutsenko, S.、Efremov, R. G.、Tsivkovskii, R. 和 Walker, J. M. (2002) 人类铜转运 ATP 酶 ATP7B（威尔逊氏病蛋白）：生化特性和调节 J. Bioenerg。生物膜。 34、351-362。