Solution NMR Studies of an ATP-Binding Cassette

描述（由申请方提供）：ATP结合盒（ABC）是ABC转运蛋白超家族的特征模块。ABC转运蛋白在所有非病毒基因组中占很大比例，并跨细胞和细胞器膜转运各种重要的货物，包括离子、氨基酸、脂质、糖和蛋白质。ABC转运蛋白与许多不同的和关键的病理学相关，如多药耐药（MDR）和囊性纤维化。ATP结合盒是ABC转运蛋白的能量产生马达结构域，但其ATP水解和机械化学能量转导的机制知之甚少。最近的晶体学研究表明，ATP诱导的ABCs的构象变化可能是ATP水解和溶质转运之间的重要耦合机制。本奖学金的建议将适用于广泛的强大的解决方案核磁共振（NMR）技术来描绘ATP诱导的构象和动态变化的ABC结构域（MJ1267）的分支氨基酸转运蛋白从超嗜热古细菌。将在各种突变体中进行类似的研究，以建立ATP诱导的构象变化，ATP催化机制和重要疾病的分子病因学之间的关系。从拟议的研究中收集到的详细结构和动态信息将拓宽我们对ATP结合盒功能的基本理解，并为基于结构的囊性纤维化药物设计提供必要的知识和工具。

DESCRIPTION (provided by applicant): The ATP-binding cassette (ABC) is the signature module of the ABC transporter superfamily. ABC transporters comprise a significant percentage of all non-viral genomes and transport a variety of important cargos, including ions, amino acids, lipids, sugars and proteins, across cell and organelle membranes. The ABC transporters are associated with a number of diverse and critical pathologies, such as multi-drug resistance (MDR) and cystic fibrosis. ATP-binding cassettes are the energy generating motor domain of the ABC transporters but their mechanism of ATP hydrolysis and mechanochemical energy transduction is poorly understood. Recent crystallographic studies suggest that ATP-induced conformational changes in ABCs are likely to be the crucial coupling mechanism between ATP hydrolysis and solute translocation. The present fellowship proposal will apply a wide array of powerful solution nuclear magnetic resonance (NMR) techniques to delineate ATP-induced conformational and dynamic changes in the ABC domain (MJ1267) of the branched amino acid transporter from a hyperthermophilic archaebacterium. Similar studies will be carried out in a variety of mutants to establish the relationship between ATP-induced conformational changes, the mechanism of ATP-catalysis and molecular etiology of important diseases. Detailed structural and dynamic information gleaned from the proposed studies will broaden our fundamental understanding of ATP-binding cassette function and contribute essential knowledge and tools to the structure-based drug design for cystic fibrosis.