Nano Accordions for Probing Biomolecular Interactions

用于探测生物分子相互作用的纳米手风琴

• 批准号: 6933923
• 负责人: ALEXANDER D LI
• 金额: $ 21.73万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933923
• 关键词: DNA binding protein; cell transformation; chemical synthesis; chromophore; conformation; fluorescence microscopy; fluorescence spectrometry; intermolecular interaction; macromolecule; molecular dynamics; molecular shape; nonhistone nucleoprotein; nucleic acid sequence; nucleic acid structure; protein binding

DESCRIPTION (provided by applicant): Protein and deoxyribonucleic acid (DNA) interactions are ubiquitous and play vital roles in controlling functional cellular systems and abnormal cell transformations (e.g., tumor progression). Upon binding, the DNA and/or proteins frequently undergo conformational changes due to the requirement of new complex formation. We hypothesize that these "molecular mechanics" can be probed with a man-made molecular accordion consisting of a string of fluorescent chromophores linked with foldable hinges. Molecular conformation changes will exert forces on the accordion, thereby tuning the fluorescent emission colors. As a first step, we will use solid phase synthesis to construct molecular accordions consisting of fluorescent chromophores and DNA sequences known to promote protein binding. The accordion-protein interactions will be carried out in vitro with wellcontrolled parameters. Our specific aims are to design and synthesize molecular accordions with multiple fluorescent chromophores on DNA and to apply fluorescent spectroscopy and microscopy in controlled conditions to gauge protein-DNA interactions. In particular, we will start with HMGA1 protein, which is a diagnostic marker for neoplastic transformation and metastatic potential of many type cancers. In this project, we plan to determine the parameters that define the scope of the formation of protein-DNA-accordion complex; these parameters will be essential for proposed future in vivo applications of molecular accordions. Our long-term objectives are to use these molecular accordions to probe regulatory events in vivo and to further our understanding of abnormal cell transformations underlying molecular mechanism of diseases.

描述（由申请人提供）：蛋白质和脱氧核糖核酸（DNA）相互作用无处不在，在控制功能性细胞系统和异常细胞转化（例如肿瘤进展）中发挥着至关重要的作用。结合后，由于新复合物形成的需要，DNA 和/或蛋白质经常发生构象变化。我们假设这些“分子力学”可以用人造分子手风琴来探测，该分子手风琴由一串与可折叠铰链连接的荧光发色团组成。分子构象的变化会对手风琴施加力，从而调整荧光发射颜色。第一步，我们将使用固相合成来构建由荧光发色团和已知促进蛋白质结合的 DNA 序列组成的分子手风琴。手风琴-蛋白质相互作用将在体外以良好控制的参数进行。我们的具体目标是设计和合成 DNA 上具有多个荧光发色团的分子手风琴，并在受控条件下应用荧光光谱和显微镜来测量蛋白质-DNA 相互作用。特别是，我们将从 HMGA1 蛋白开始，它是许多类型癌症的肿瘤转化和转移潜力的诊断标志物。在这个项目中，我们计划确定定义蛋白质-DNA-手风琴复合物形成范围的参数；这些参数对于分子手风琴未来的体内应用至关重要。我们的长期目标是利用这些分子手风琴来探测体内的调控事件，并进一步了解疾病分子机制背后的异常细胞转化。