Mechanistic Investigations of Ketosteroid Isomserase

酶是细胞内的分子，它使生命所必需的化学反应快速而正确地发生。这些反应控制着细胞过程的各个方面，从从食物中提取能量，到构建细胞和器官所需的结构，再到生物体经历不同环境和压力时对这些过程的精细控制。本研究旨在通过研究一种高度可处理的模式酶，细菌酮类固醇异构酶，来促进对酶作用的基本理解。这种酶有几个特点，将使其研究在前所未有的深度和广度。具体来说，这项研究需要精细的结构，动态和生化研究的混合。此外，这种酶易于化学合成，因此可以系统而彻底地引入单个氨基酸水平的变化，而不受20种天然存在的氨基酸的限制。最广泛地说，酶机制中一个悬而未决的核心问题是催化发生的活性位点周围的蛋白质结构的作用，这项研究将为酶环境的性质提供深刻的测试。了解酶的功能是了解生命本身的基础。此外，大多数药物靶点都是酶，因此对酶作用的更深入的基本理解可能使研究人员能够更好地设计出有效的抑制剂，这些抑制剂可以作为多种疾病状态的药物。对酶作用的深入了解也可以提供设计新酶所需的信息，这些酶可以催化新的有益的反应，这些反应可以用于制药和化学工业。该项目的更广泛影响包括培养生物化学严谨、定量方面的研究生，这一背景将使这些学生在其个人职业生涯、教师、应用或基础研究以及从科学角度评估政策方面以多种方式作出贡献。此外，研究的基本性质使得本科生有可能获得有意义的研究经验。特别是，有计划确保将代表性不足的少数民族纳入这项研究，这一机会可能极大地有助于促进这些学生在未来融入科学界。

Enzymes are the molecules within cells that make the chemical reactions necessary for life happen fast and happen correctly. These reactions govern all aspects of cellular processes, from the extraction of energy from food, to the building of structures needed for constructing cells and organs, to the exquisite control of these processes as an organism experiences different environments and stresses. This research aims tocontribute to the fundamental understanding of enzyme action via investigation of a highly tractable, model enzyme, bacterial ketosteroid isomerase. This enzyme has several features that will allow its study at unprecedented depth and breadth. Specifically, this research entails a finely crafted blend of structural, dynamic and biochemical studies. Furthermore, this enzyme is tractable for chemical synthesis so that changes at the level of individual amino acids can be introduced systematically and thoroughly, without the limitations of the 20 naturally occurring amino acids. Most broadly, a central unanswered question in enzyme mechanism is the role of the protein structure surrounding the active site where catalysis takes place, and this research will provide incisive tests of the properties of the enzyme environment.Understanding how enzymes function is fundamental for understanding life itself. Furthermore, the majority of drug targets are enzymes, so that a deeper fundamental understanding of the action of enzymes may allow researchers to better design potent inhibitors that can act as drugs for numerous disease states. A deep understanding of enzyme action may also provide the information needed to design new enzymes thatcatalyze new and beneficial reactions that can be used in the pharmaceutical and chemical industries.The broader impact of this project involves the training of graduate students in rigorous, quantitative aspects of biological chemistry, a background that will allow these students to contribute in many ways in their individual careers, as teachers, in applied or fundamental research, and in evaluating policy from a scientific standpoint. Further, the basic nature of the research renders it possible for undergraduates to obtain meaningful research experiences. In particular there are plans to ensure the inclusion of underrepresented minorities in this research, an opportunity that may greatly help stimulate the inclusion of such students in the future in the scientific community.