PECASE: A Difference Infrared Spectroscopic Study of a Nucleotide Binding Protein

PECASE：核苷酸结合蛋白的差异红外光谱研究

• 批准号: 9733566
• 负责人: Gina MacDonald
• 金额: $ 50万
• 依托单位: James Madison University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9733566&HistoricalAwards=false
• 关键词: PECASE; Difference; Infrared; Spectroscopic; Study

9733566 MacDonald This CAREER plan integrates research and educational components and involves undergraduate chemistry and biology majors as well as high school teachers in all aspects of the project. The goal of the research is to gain further insight into the regulations and mechanism of a protein involved in DNA repair and recombination. Fourier transform infrared (FTIR) difference spectroscopy will be used to investigate the relationship between protein structure and function in the Escherichia coli protein, RecA. These studies should result in a greater understanding of all allosteric changes in RecA that are responsible for regulating DNA strand exchange, a process crucial to DNA recombination and repair. RecA function is regulated by nucleotide binding. Initial studies will focus on the binding of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) to RecA. Vibrational spectroscopy will be used to identify specific changes in protein structure induced by the binding of ADP (RecA conformation with decreased affinity for DNA), and ATP (RecA conformation with a higher affinity for DNA). These studies will be performed under a variety of conditions in order to investigate how nucleotide binding affects protein structure in both the active and inactive RecA nucleoprotein filaments. These studies will provide the groundwork for future RecA studies such as (1) how the previously studied changes in RecA structure are affected by other proteins known to interact with RecA in vitro and (2) time-resolved studies that follow nucleotide binding and strand exchange in real time. High school teachers, as well as James Madison University undergraduates in chemistry and biology ,and secondary education minors, will play instrumental roles in both the educational and research aspects of this CAREER grant. A larger number of participants can be involved in research efforts by providing two types of projects (1) biochemical and difference infrared studies of RecA (2) research effor ts that utilize departmental equipment and can be incorporated into the new project oriented biochemistry laboratory. Students involved will gain valuable research experience, learn current biochemical and spectroscopic techniques, and have a greater understanding of the scientific method. The undergraduate participants will contribute to a greater understanding of structure-function relationships in RecA and an exciting new biochemical laboratory. Including current and future high school teachers in this project will ultimately help to attract the next generation of scientists.

这个职业规划整合了研究和教育的组成部分，涉及本科化学和生物专业以及高中教师在项目的各个方面。这项研究的目标是进一步了解参与DNA修复和重组的蛋白质的调节和机制。傅里叶变换红外（FTIR）差分光谱将用于研究大肠杆菌蛋白RecA的蛋白质结构与功能之间的关系。这些研究将有助于更好地了解RecA中所有负责调节DNA链交换的变构变化，这是DNA重组和修复的关键过程。RecA功能受核苷酸结合调控。最初的研究将集中在三磷酸腺苷（ATP）和二磷酸腺苷（ADP）与RecA的结合上。振动光谱学将用于鉴定由ADP（对DNA亲和力降低的RecA构象）和ATP（对DNA亲和力较高的RecA构象）结合引起的蛋白质结构的特定变化。这些研究将在各种条件下进行，以研究核苷酸结合如何影响活性和非活性RecA核蛋白细丝中的蛋白质结构。这些研究将为未来的RecA研究提供基础，例如：(1)先前研究的RecA结构变化如何受到已知与RecA在体外相互作用的其他蛋白质的影响；(2)实时跟踪核苷酸结合和链交换的时间分辨率研究。高中教师，以及詹姆斯麦迪逊大学化学和生物学专业的本科生，以及中学教育的未成年人，将在这项职业资助的教育和研究方面发挥重要作用。通过提供两种类型的项目(1)RecA的生化和差异红外研究(2)利用部门设备的研究工作，可以纳入新的面向项目的生物化学实验室。参与的学生将获得宝贵的研究经验，学习当前的生化和光谱技术，并对科学方法有更深入的了解。本科学生将有助于更好地理解RecA和令人兴奋的新生化实验室的结构-功能关系。在这个项目中包括现在和未来的高中教师将最终有助于吸引下一代科学家。