CAREER: Simultaneous Single-Molecule Manipulation and Visualization for Study of ATP Coupling to Base Pair Unzipping

在化学系化学测量和成像计划的支持下，密歇根大学的程伟教授和他的团队将开发一种超灵敏技术，允许一次一个分子地操纵和可视化生化反应。他们将使用这项技术来研究在解旋酶催化的反应中，ATP是如何被用于碱基对解压的。2%的真核基因组编码解旋酶蛋白。这些蛋白质无处不在，它们使用三磷酸腺苷来解压DNA或RNA，这样遗传信息就可以进行进一步的处理。然而，这些酶如何利用三磷酸腺苷进行BP解压的机制仍然知之甚少。该项目专注于开发一种单分子技术来同时操纵和可视化丙型肝炎病毒NS3蛋白的反应，NS3蛋白是一种模型解旋酶。这项技术的成功可以使我们了解解旋酶机制的基础知识，并允许对广泛的生化反应进行精确的定量测量。该项目的更广泛影响涉及研究和教学活动，旨在强调技术创新在科学进步中的重要性，特别是对生物化学和生物物理感兴趣的学生。程教授建议教授生物化学课程，重点是使重大科学发现成为可能的技术创新。此外，他计划在他的实验室里建立一个年度单分子展示，向当地的高中生和大学生开放。他希望通过简单的演示实验和动手体验，激发学生对基础科学的兴趣。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Wei Cheng at University of Michigan and his group will develop an ultrasensitive technique that allows manipulation and visualization of a biochemical reaction, one molecule at a time. They will use this technique to study how ATP is consumed for base pair unzipping in a helicase-catalyzed reaction. 2% of the eukaryotic genome encodes helicase proteins. These proteins are ubiquitous and use ATP to unzip DNA or RNA so that genetic information is accessible for further processing. However, the mechanism of how these enzymes use ATP for bp unzipping remains poorly understood. This project focuses on the development of a single-molecule technique to simultaneously manipulate and visualize the reaction of hepatitis C virus NS3 protein, a model helicase. The success of this technique can lead to fundamental knowledge in our understanding of helicase mechanism, and allows quantitative measurement of a broad range of biochemical reactions in exquisite detail.The broader impacts of this project involve research and teaching activities that aim to emphasize the importance of technical innovation in advancement of science, particularly to students with interests in biochemistry and biophysics. Professor Cheng proposes to teach biochemistry classes with emphasis on technical innovation that enabled important scientific discoveries. Moreover, he plans to establish an annual single-molecule showcase in his lab, which will be open to local high school and college students. Through simple demonstration experiments and hands-on experience, he hopes to inspire the students' interest in basic science.