Thermodynamics of Coupled Binding of Zn(II) and DNA to a Zinc Finger Tumor Suppre

Zn(II) 和 DNA 与锌指肿瘤抑制物偶联结合的热力学

• 批准号: 7761972
• 负责人: Brian R. Gibney
• 金额: $ 8.54万
• 依托单位: BROOKLYN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7761972
• 关键词: Affinity; Amyotrophic Lateral Sclerosis; Binding; Binding Proteins; Biological Assay; Biological Models; Burial; Circular Dichroism; Clinical Trials; Commit; Complex; Coupled; DNA; DNA Binding; DNA-Binding Proteins; Data; Development; Diabetic Neuropathies; Dissociation; Drug Design; Equilibrium; Fluorescence Spectroscopy; Funding; Future; Gene Activation; Genes; Genetic Transcription; Genome; Goals; Healthcare; Heart Diseases; Human; Human Genome; In Vitro; Ions; Kinetics; Lead; Malignant Neoplasms; Mammalian Cell; Measures; Metal Ion Binding; Metalloproteins; Metals; NMR Spectroscopy; Nephroblastoma; Oncogenes; Peptides; Pharmaceutical Preparations; Positioning Attribute; Process; Protein Binding; Proteins; Publications; RNA; Research; Site; Social Welfare; Solvents; Structure; Testing; Therapeutic; Thermodynamics; Transcriptional Regulation; Tryptophan; Tumor Suppressor Proteins; Work; Zinc; Zinc Fingers; base; college; combinatorial; cost; design; drug candidate; gene therapy; human disease; improved; insight; novel therapeutics; programs; protein expression; protein folding; public health relevance; success; tool; transcription factor; tumor

DESCRIPTION (provided by applicant): The long-term goal of this research is to improve the rational design of artificial zinc finger protein (ZFP) transcription factor therapeutics by characterizing the fundamental thermodynamics of ZFP transcription factor interactions with zinc metal-ions and DNA. ZFPs, the largest single class of metalloproteins in the human genome, regulate gene transcription and, therefore, protein expression, by binding both zinc metal-ions and DNA or RNA. Since ZFP transcription factors can be potentially designed to activate or repress any gene in the genome, these proteins are excellent targets for drug design. Indeed the rational design and combinatorial selection of artificial ZFPs is beginning to yield potential drug candidates, one of which is in clinical trials for diabetic neuropathy and Lou Gehrig's disease. Furthermore, recent success in ZFP protein transduction in mammalian cells indicates they may be suitable for gene therapy applications. One issue that slows the progress of artificial ZFP design is coupled energetics of zinc metal-ion binding, DNA-binding, and protein folding in ZFPs. Based on our success in parsing apart the energetics of zinc metal-ion binding and protein folding using simple designed peptides, we are in a unique position to decouple the binding of zinc metal-ions and DNA in a natural ZFP, the Wilms tumor suppressor. We will focus our research efforts in delineating the fundamental thermodynamics of ZFP binding to zinc metal-ions and DNA. Our approach is to measure the equilibrium thermodynamics of Zn(II) and DNA binding to ZFPs. Based on the current lack of understanding of the coupled binding of Zn(II) and DNA to ZFPs, we have identified two Specific Aims to be completed during the requested funding period. Specific Aim 1: Study of zinc finger protein Zn(II) affinity in the presence of DNA. Specific Aim 2: Study of apo- zinc finger protein folding. PUBLIC HEALTH RELEVANCE: The detailed understanding of the interrelationship of Zn(II) and DNA binding to zinc finger proteins provided by this study will lead to a greater understanding of general metalloregulatory processes, and specifically gene activation in human cancer. These results will facilitate both the rational design of tumor suppressor drugs and improved prediction of the process of cancer gene activation. Therefore, benefits to healthcare and the general welfare of the public are anticipated from the successful completion of this work.

描述（由申请人提供）：本研究的长期目标是通过表征ZFP转录因子与锌金属离子和DNA相互作用的基本热力学来改进人工锌指蛋白（ZFP）转录因子治疗剂的合理设计。ZFP是人类基因组中最大的一类金属蛋白，通过结合锌金属离子和DNA或RNA来调节基因转录，从而调节蛋白质表达。由于ZFP转录因子可以被潜在地设计成激活或抑制基因组中的任何基因，因此这些蛋白质是药物设计的极好靶点。事实上，人工ZFP的合理设计和组合选择开始产生潜在的候选药物，其中之一是在糖尿病神经病变和卢伽雷病的临床试验。此外，最近在哺乳动物细胞中ZFP蛋白转导的成功表明它们可能适合于基因治疗应用。减缓人工ZFP设计进展的一个问题是ZFP中锌金属离子结合、DNA结合和蛋白质折叠的耦合能量学。基于我们使用简单设计的肽成功地解析了锌金属离子结合和蛋白质折叠的能量学，我们处于独特的位置，以解耦锌金属离子和DNA在天然ZFP中的结合，Wilms肿瘤抑制剂。我们将集中我们的研究工作在描绘的基本热力学ZFP结合锌金属离子和DNA。我们的方法是测量Zn（II）和DNA与ZFP结合的平衡热力学。基于目前对Zn（II）和DNA与ZFP的偶联结合缺乏了解，我们确定了两个特定目标，将在申请的资助期间完成。具体目的1：研究锌指蛋白Zn（II）在DNA存在下的亲和力。具体目的2：研究载脂蛋白锌指蛋白的折叠. 公共卫生相关性：本研究提供的锌（II）和DNA结合锌指蛋白的相互关系的详细了解，将导致更好地了解一般金属调节过程，特别是在人类癌症的基因激活。这些结果将有助于肿瘤抑制药物的合理设计和改善对癌症基因激活过程的预测。因此，预计这项工作的成功完成将给医疗保健和公众的普遍福利带来好处。