RUI: Mechanistic Studies of the Firefly Luciferase Catalyzed Reactions

该项目的目标是增加对生物体如何将化学能转化为光的理解。 生物发光，即生物体发光，是一种美丽的自然现象，它让孩子们着迷，挑战了那些试图理解它的人，并为重要的研究工具提供了基础。 生物发光的例子可以在自然界的细菌、蘑菇、水母、蚯蚓、蛤、鱼和甲虫中找到。 该项目解决了萤火虫荧光素酶的结构及其在催化两个顺序反应中的详细作用之间的关系，最终导致北美萤火虫Photinus pyralis产生黄绿光。 将通过光谱和动力学方法评估通过重组DNA技术制备的新的酶，以检查突变对每个反应的影响。 这种方法也可以提供证据的荧光素酶结构中的基本构象变化的作用。 这项研究的结果将增强对萤火虫生物发光的基本理解，以及生物体将化学能转化为光的基本过程。 该项目旨在满足生产新型荧光素酶蛋白的需求，以改善目前生物发光的研究应用，并推动新产品的开发。 这项研究将在一所文科学院进行，该学院对本科生进行了有效的研究培训。 现代化的设施和设备为学生和教师提供了一个良好的环境，进行合作研究。 主要研究者积极指导本科生研究项目32年。 大约有85名本科生，其中大多数是妇女和少数民族，与他一起工作，估计70%的参与者已经进入研究生或专业学校。 这个项目的目的是提供一个有意义的研究经验，在多元文化的环境中，一个不同的学生群体。 该项目的所有学生和专业参与者将参与现代主流生物发光研究，并将为增加准备充分的毕业生进入研究生课程和专业科学职业的人数做出积极贡献。 这项研究的结果可能会导致萤火虫荧光素酶用途的改进，例如，在检测被细菌污染的食品或作为基因表达和调控的报告者。

The goal of this project is to increase the understanding of how living organisms convert chemical energy into light. Bioluminescence, the emission of light by living organisms, is a beautiful natural phenomenon that has enchanted children, challenged those who have tried to understand it, and provided the basis for an important research tool. Examples of bioluminescence can be found throughout nature in bacteria, mushrooms, jellyfish, earthworms, clams, fish and beetles. This project addresses the relationship between the structure of the enzyme firefly luciferase and its detailed role in the catalysis of two sequential reactions that ultimately lead to the production of yellow-green light in the North American firefly, Photinus pyralis. New luciferases made by recombinant DNA techniques will be evaluated by spectroscopic and kinetics methods to examine the effects of mutations on each reaction. This approach may also provide evidence of the role of essential conformational changes in the luciferase structure. The results of this research will enhance the basic understanding of firefly bioluminescence and the fundamental process by which living organisms convert chemical energy into light. The project addresses the need to produce novel luciferase proteins to improve current research applications of bioluminescence and to advance the development of new ones.Broader Impacts. This research will be carried out at a liberal arts college with an established record of effective research training of undergraduate students. Modern facilities and equipment provide an excellent environment for students and faculty to engage in collaborative research. The principal investigator has actively directed undergraduate research projects for 32 years. Approximately 85 undergraduate students, the majority of whom are women and minorities, have worked with him, and an estimated 70% of these participants have entered graduate or professional school. The aim of this project is to offer a meaningful research experience to a diverse group of students in a multicultural environment. All student and professional participants in this project will be involved in modern mainstream bioluminescence research and will contribute positively to increasing the numbers of well-prepared graduates for entry into graduate programs and professional scientific careers. Results of this research may lead to improvements in the uses of firefly luciferase, for example, in the detection of food contaminated with bacteria or as a reporter of gene expression and regulation.