Fluorous Proteins: Structure, Stability, and Biological Activity

氟蛋白：结构、稳定性和生物活性

• 批准号: 0640934
• 负责人: E. Neil Marsh
• 金额: $ 43.5万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2010-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0640934&HistoricalAwards=false
• 关键词: Fluorous; Proteins; Structure; Stability; Biological

With this award, the Organic and Macromolecular Chemistry Program supports Neil Marsh and Hashim M. Al-Hashimi both of the University of Michigan whose research will advance the area of protein design by engineering some of the novel properties of fluorocarbons into biological molecules. This will be achieved by synthesizing proteins that contain extensively fluorinated ('fluorous') analogs of hydrophobic amino acids in their hydrophobic cores. Fluorous amino acids are predicted to stabilize proteins against unfolding by heat and organic solvents and to facilitate protein: protein recognition through specific fluorocarbon-fluorocarbon interactions. Fluorinated versions of a dimeric RNA-binding protein, Rop, will be synthesized in which the hydrophobic core of Rop will be repacked with the fluorous analog of leucine, hexafluoroleucine. This protein is small enough (63 residues) to be efficiently synthesized by peptide synthesis, which will allow fluorous amino acids to be introduced site specifically. Rop protein has been extensively used as a model system for investigating protein stability and folding, and as a template for protein re-design. These data will serve as a useful reference for the present study. A variety of physical techniques (such as circular dichroism, microcalorimetry and analytical ultracentrifugation) will be used to investigate the effect of fluorination on the biological activity, structure and stability of fluorous Rop proteins. An important innovation will be the use of residual dipolar coupling (RDC) NMR measurements to perform detailed comparisons of the effect of fluorination on the structure and conformational rigidity of the protein. The experiments will address fundamental questions about the impact of fluorination on protein structure and dynamics.This award from the Organic and Macromolecular Chemistry Program supports Professors Neil Marsh and Hashim M. Al-Hashimi both of the University of Michigan whose research will impact attempts to design biosensors and enzymes used in industrial processes, where stability towards extremes of temperature and pH and towards organic solvents is necessary. There is the potential for fluorous proteins to find uses in medical imaging by exploiting the high NMR sensitivity of Fluorine 19 or their enhanced biological stability could lead to uses as therapeutic agents or vehicles for drug delivery. The project will advance the education, training and professional development of undergraduates, graduate students and postdoctoral scientists in the inter-disciplinary area of chemical biology and biophysics. To broaden their education further, a joint interdisciplinary group meeting and journal club will be initiated. Their professional development will be enhanced by active participation in the dissemination of their results, both through drafting manuscripts and progress reports, and through oral and poster presentations at local and national scientific meetings.

有了这个奖项，有机和高分子化学计划支持尼尔马什和哈希姆M。他们的研究将通过将碳氟化合物的一些新特性改造成生物分子来推进蛋白质设计领域。这将通过合成在其疏水核心中含有疏水氨基酸的广泛氟化（“氟”）类似物的蛋白质来实现。预计含氟氨基酸可稳定蛋白质，使其免受热和有机溶剂的解折叠，并通过特定的碳氟化合物-碳氟化合物相互作用促进蛋白质：蛋白质识别。将合成二聚体RNA结合蛋白Rop的氟化版本，其中Rop的疏水核心将用亮氨酸的氟类似物六氟亮氨酸重新包装。这种蛋白质足够小（63个残基）以通过肽合成有效地合成，这将允许氟氨基酸被位点特异性地引入。ROP蛋白已被广泛用作研究蛋白质稳定性和折叠的模型系统，并作为蛋白质重新设计的模板。这些数据将作为本研究的有益参考。各种物理技术（如圆二色性，微量热和分析超离心）将被用来研究的氟Rop蛋白的生物活性，结构和稳定性的影响。一个重要的创新将是使用残余偶极耦合（RDC）NMR测量，以执行详细的比较的影响，对蛋白质的结构和构象刚性的双金属离子。这些实验将解决有关氟化对蛋白质结构和动力学影响的基本问题。有机和大分子化学项目的这一奖项支持Neil Marsh教授和Hashim M.密歇根大学的Al-Hashimi博士的研究将影响设计用于工业过程的生物传感器和酶的尝试，其中对极端温度和pH值以及有机溶剂的稳定性是必要的。通过利用氟19的高NMR灵敏度，氟蛋白有可能在医学成像中找到用途，或者它们增强的生物稳定性可能导致用作药物递送的治疗剂或载体。该项目将促进化学生物学和生物物理学跨学科领域本科生、研究生和博士后科学家的教育、培训和专业发展。为了进一步扩大他们的教育，将发起一个联合跨学科小组会议和期刊俱乐部。他们的专业发展将通过积极参与传播其成果得到加强，包括起草手稿和进度报告，以及在地方和国家科学会议上作口头和海报介绍。