CAREER: Development of Gas-phase, Post-ionization Structural Tools for the Study of DNA-protein Complexes

在化学系化学测量和成像计划的支持下，以及分子和细胞生物科学系分子生物物理组的共同资助下，佛罗里达国际大学的Francisco Fernandez-Lima教授和他的团队正在开发集成结构生物学技术的新分析工具，以表征对理解生物和生化过程至关重要的DNA-蛋白质相互作用。许多重要的生化和生物系统不适用于传统的分析方法(例如磁共振或X射线结晶学)，这就需要新的和补充的方法来更好地表征生物分子和生物分子化合物。费尔南德斯-利马博士基于质谱学(MS)的方法显著增强了表征能力。在这些研究工作的同时，他计划采取一种多层次的拓展方法，包括一系列研讨会、本科生和研究生课程、博物馆开放活动、亲身体验MS、暑期实习以及开发虚拟MS实验系列。该计划与当地的K-12学校、邻近的社区大学和FIU的学生培训项目合作，旨在加强学习和接触现代MS技术，并招收来自少数族裔的学生进入科学、技术、工程和数学领域。通过整合质谱学(MS)、俘获离子迁移率谱(TIMS)、氢/氘背交换(HDX)和离子光谱技术，费尔南德斯-利马教授和他的团队正在研究内在无序的系统，重点是DNA结合蛋白质和DNA蛋白质复合体。这种新的实验方法有可能改变我们询问生物分子的方式，提供关于构象系综中共存状态(例如，平衡中间体和动力学中间体)的详细、有用的结构信息。实验以新的结构信息为目标，使用迁移率选择的构象询问来研究从溶液到气相的构象变化，并提供关于构象重排的能垒及其时间尺度的信息。该项目的研究成果可能会影响我们对驱动生物分子和生物分子复合体折叠和去折叠的结构基序的基本理解。此外，佛罗里达国际大学还建立了一个外展教育项目“FIUMASS：MS Experience for All”，以增加研究生、本科生和高中生对结构质谱学实践方面的了解。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry and co-funding from the Molecular Biophysics Cluster of the Division of Molecular and Cellular Biosciences, Professor Francisco Fernandez-Lima and his group at Florida International University are developing new analysis tools integrating structural biology techniques to characterize DNA-protein interactions critical to understanding biological and biochemical processes. Many important biochemical and biological systems are not amenable to traditional analytical approaches (e.g., magnetic resonance or X-ray crystallography), creating a need for new and complementary methods to better characterize biomolecules and biomolecular complexes. Dr. Fernandez-Lima's mass spectrometry (MS)-based approaches offer significant enhancement of characterization capabilities. In parallel with these research efforts, he plans a multi-level outreach approach integrating a seminar series, undergraduate and graduate courses, museum open-house events, hands-on MS experiences, summer internships, and development of a virtual MS experiment series. Partnering with local K-12 schools, neighboring community colleges, and student training programs at FIU, this plan aims to enhance learning and exposure to modern MS techniques and to recruit students from underrepresented minorities into science, technology, engineering and mathematics.By integrating mass spectrometry (MS), trapped ion mobility spectrometry (TIMS), hydrogen/deuterium back-exchange (HDX) and ion spectroscopy techniques, Prof. Fernandez-Lima and his group are studying intrinsically disordered systems, with a focus on DNA-binding proteins and DNA-protein complexes. This new experimental approach has the potential to transform the way we interrogate biological molecules, providing detailed, useful structural information on co-existing states in the conformational ensemble (e.g., equilibrium and kinetic intermediates). Experiments target new structural information, employing mobility-selected conformation interrogation to enable study of conformational changes from solution to the gas-phase, and providing information on the energy barriers for conformational rearrangement and their time scales. The research outcomes of this project may impact our fundamental understanding of the structural motifs that drive the folding and unfolding of biomolecules and biomolecular complexes. In addition, an outreach educational program "FIUMASS: MS Experience for All" has been established at Florida International University to increase the exposure of graduate, undergraduate, and high school minority students to the practical aspects of structural mass spectrometry.