RUI: Mechanistic Studies of the Firefly Luciferase Catalyzed Reactions

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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.