RUI: Mutagenesis and Mechanistic Studies of the Reactions Catalyzed by Firefly Luciferase

北美萤火虫石楠的光发射是人们熟悉的生物发光的例子，生物发光是由活的有机体发出的光。生物将化学能转化为光的这一基本过程具有许多潜在的实验室和医学应用，包括使用萤火虫荧光素酶作为基因表达和调控的报告。这项研究将讨论萤火虫荧光素酶的结构与其在两个导致发光的连续反应中的催化作用之间的关系。突变的荧光素酶将通过重组DNA技术来制造，并进行评估，以检查突变对每个反应的影响。这种方法也可能为荧光素酶结构中基本重要构象变化的发生和作用提供证据。该项目的结果将加强对萤火虫生物发光以及更大的腺苷形成酶超家族的基本理解，该超家族包括萤火虫荧光素酶和许多非生物发光蛋白。更广泛的影响：这项研究将在康涅狄格学院进行，这是一所文科学院，在对本科生进行有效的研究培训方面有着既定的记录。学生将与教师进行合作研究，那些对该项目做出实质性贡献的人将成为手稿的共同作者，并在科学会议上展示他们的结果。所有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生涯做出积极贡献。

Light emission by the North American firefly Photinus pyralis is a familiar example of bioluminescence, the emission of light by a living organism. This fundamental process by which living organisms convert chemical energy into light has many potential laboratory and medical applications, including the use of firefly luciferase as a reporter of gene expression and regulation. This research will address the relationship between the structure of firefly luciferase and its role in the catalysis of two sequential reactions that result in the emission of light. Mutant luciferases will be made by recombinant DNA techniques and evaluated to examine the effects of mutations on each reaction. This approach may also provide evidence to the occurrence and role of fundamentally important conformational changes in the luciferase structure. The results of this project should enhance the basic understanding of firefly bioluminescence, as well as the larger adenylate-forming enzyme superfamily, which includes firefly luciferase and many non-bioluminescent proteins. Broader Impacts: This research will be carried out at Connecticut College, a liberal arts college with an established record of effective research training of undergraduate students. Students will engage in collaborative research with faculty and those who make substantive contributions to the project will become co-authors of manuscripts and present their results at scientific meetings. All participants 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.