CAREER: Mechanistic Studies of Tyrosine Derived Cofactors

铜胺氧化酶(CAO)和赖氨酸氧化酶(LOX)是分别由酪氨酸、2，4，5-三羟基苯丙氨酸醌(TPQ)和赖氨酸酪氨酸醌(LTQ)翻译后形成的含醌辅因子的蛋白质家族。这些酶催化生物胺的修饰，并在细胞信号和胶原形成中发挥作用。对这些苯醌辅因子的结构/功能的研究对于表征它们的化学性质的差异是至关重要的。尽管TPQ在CAO催化中的化学作用已经得到了很好的研究，但对LTQ的研究还很少。此外，铜在这两种酶中的催化作用仍不清楚。该研究项目将进一步明确TPQ和LTQ的化学性质以及铜在CAO催化中的作用，并开发一个模型系统来了解LTQ在LOX中的化学作用。本研究综合运用了生物化学、分子生物学、生物无机化学和合成有机化学等工具和技术。这项研究的结果不仅将拓宽对藜麦蛋白的理解，还将影响对自然界中广泛存在的其他交联醌的基本理解。这将有助于理解在生物系统中选择一个醌辅助因子而不是另一个的进化优势。更广泛的影响：这个多学科项目为研究生和本科生提供了一个在化学和生物学的交界处学习和参与国际合作的绝佳机会。将启动一项外展计划，邀请当地高中的女学生在堪萨斯大学化学系呆两周。这些学生将在实验室工作，并与女性教职员工和研究生会面，讨论科学和女性在化学领域面临的问题。这些经历的目的是鼓励更多的妇女追随化学职业，从一开始就表明学术环境可以支持妇女。研讨会还将表明，化学是一门多样化的学科，其范围比通常在高中教授的要广泛得多。同时，将开发一门针对高年级本科生和研究生的化学生物学课程，以演示如何使用化学方法来理解生物系统的基本方面。这门课程将包括蛋白质化学，特别是翻译后修饰以及尖端发现的概述，旨在向学生展示化学家可以在生物科学的尖端做出重要贡献。参加堪萨斯大学少数民族学生发展倡议(IMSD)-生物科学倡议项目的学生将被鼓励报名参加这门课程。首席调查员将通过这个项目积极招募少数民族本科生进入她的实验室，帮助他们在化学生物学领域进行研究生学习和研究生涯。

Copper amine oxidase (CAO) and lysyl oxidase (LOX) belong to a family of proteins that contain quinone cofactors that are formed post-translationally from tyrosine, 2,4,5-trihydroxy-phenylalanine quinone (TPQ), and lysine tyrosylquinone (LTQ), respectively. These enzymes catalyze the modification of biogenic amines and play roles in cell signaling and collagen formation. The structure/function study of these quinone cofactors is essential to characterize the differences in their chemical properties. Although the chemical function of TPQ in CAO catalysis has been well studied, very little is known about LTQ. Further, the role of copper in catalysis in both enzymes remains unclear. The research project will further define the chemical properties of TPQ and LTQ and the role of copper in CAO catalysis, as well as develop a model system to understand the chemical function of LTQ in LOX. The research utilizes the tools and techniques of biochemistry, molecular biology, bioinorganic and synthetic organic chemistry. The results of this research will not only broaden understanding of quinoproteins but also impact fundamental understanding of other cross-linked quinones that are widely found in nature. This will aid in understanding the evolutionary advantage of selecting one quinone cofactor over the other in biological systems.Broader Impact: This multidisciplinary project provides an excellent opportunity for graduate and undergraduate students to study at the interface of chemistry and biology and participate in an international collaboration. An outreach program will be initiated to invite female students from local high schools to spend two weeks in the Chemistry department at the University of Kansas. The students will work in the laboratory and meet with female faculty and graduate students to discuss both science and the issues facing women in chemistry. These experiences are intended to encourage more women to follow careers in chemistry by demonstrating at the outset that the academic environment can be supportive to women. The workshop will also show that chemistry is a diverse subject that is much broader in scope than what would typically be taught in high school. In concert, a chemical biology course targeting upper level undergraduate and graduate students will be developed to demonstrate how chemical methods are used to understand fundamental aspects of biological systems. The course will include protein chemistry, especially post-translational modifications as well as a survey of cutting edge discoveries, intended to show students that chemists can make important contributions at the cutting-edge of biological science. Students participating in the Initiative for Minority Student Development (IMSD)-Bioscience Initiative program at the University of Kansas will be encouraged to enroll in this course. The principal investigator will actively recruit minority undergraduate students through this program into her laboratory to help them in pursuing graduate studies and a research career in the field of chemical biology.