STRUCTURAL ANALYSIS OF WILSON DISEASE ATPASE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 铜是人体呼吸、神经元功能、结缔组织形成、内分泌过程和自由基解毒等多种酶活性所必需的。威尔逊病ATPase和门克斯病ATPase调节细胞中铜的浓度，并将铜输送到生物合成途径。这些蛋白质是许多突变的目标，这些突变会导致严重的代谢紊乱。重要的是，威尔逊和门克斯ATPase参与了癌症对铂类化疗药物的耐药性。为了了解铜ATPase活性周期中的关键步骤，我们将使用多维核磁共振(NMR)来研究Wilson和Menkes蛋白质的孤立结构域或功能模块的结构、分子运动和相互作用。多维核磁共振是一种专门适合研究蛋白质在溶液中的结构和动力学的技术。我们将追踪铜转运ATPase与底物结合的分子事件序列，并分析几种常见致病突变的结构基础。为了追踪铜在肝豆状核变性和门克斯病ATPase中的途径，我们将尝试定位细胞膜上的铜结合部位。这项工作有望提高对一类重要转运蛋白的认识，并为铜代谢紊乱的分子基础提供新的见解。威尔逊和门克斯ATPase各个结构域的结构将有助于设计这些酶的新抑制剂和调节剂，这可能有助于克服癌症中某些类型的耐药性。

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. Copper is required for the activity of many enzymes involved in respiration, neuron function, formation of connective tissue, endocrine processes, and radical detoxification in the human body. Wilson disease ATPase and Menkes disease ATPase regulate copper concentration in the cell and deliver copper to biosynthetic pathways. These proteins are targets of many mutations that cause severe metabolic disorders. Importantly, Wilson and Menkes ATPases are involved in cancer resistance to platinum- based chemotherapeutic drugs. To understand the critical steps in the activity cycle of the copper ATPases, we will investigate the structure, molecular motions and interactions of the isolated domains, or functional modules, of the Wilson and Menkes proteins using multidimensional Nuclear Magnetic Resonance (NMR), a technique uniquely suited for studying protein structure and dynamics in solution. We will trace the sequence of molecular events involved in substrate binding by copper-transporting ATPases and analyze the structural basis of several frequent disease causing mutations. To trace the pathway of copper in Wilson and Menkes disease ATPases, we will attempt to map the copper-binding site in the cell membrane. This work is expected to improve the understanding of an important class of transport proteins and provide a new insight into the molecular basis of the disorders of copper metabolism. Structures of the individual domains of Wilson and Menkes ATPase will facilitate design of the new inhibitors and modulators of these enzymes, which may help to overcome certain types of drug resistance in cancer.