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项目的支持下，并在分子和细胞生物科学系分子生物物理学小组的共同资助下，弗朗西斯科·费尔南德斯-利马教授和他在佛罗里达国际大学的研究小组正在开发新的分析工具，将结构生物学技术整合在一起，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。许多重要的生物化学和生物系统不适合传统的分析方法（例如，磁共振或X射线晶体学），产生了对新的和互补的方法的需要，以更好地表征生物分子和生物分子复合物。Fernando-Lima博士基于质谱（MS）的方法显著增强了表征能力。 在这些研究工作的同时，他计划采用多层次的外展方法，将研讨会系列，本科生和研究生课程，博物馆开放日活动，实践MS经验，暑期实习以及虚拟MS实验系列的开发整合在一起。该计划与当地K-12学校、邻近的社区学院和FIU的学生培训项目合作，旨在加强对现代MS技术的学习和接触，并招募来自代表性不足的少数民族的学生进入科学、技术、工程和数学领域。通过整合质谱（MS）、捕获离子迁移谱（TIMS）、氢/氘反交换（HDX）和离子光谱技术，Fernando-Lima教授和他的团队正在研究本质上无序的系统，重点是DNA结合蛋白和DNA-蛋白质复合物。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的详细、有用的结构信息（例如，平衡和动力学中间体）。 实验针对新的结构信息，采用流动性选择的构象询问，使研究从溶液到气相的构象变化，并提供信息的能量障碍构象重排及其时间尺度。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物折叠和展开的结构基序的基本理解。此外，在佛罗里达国际大学建立了一个推广教育方案“FIUMASS：MS经验为所有人”，以增加研究生、本科生和高中少数民族学生对结构质谱法实践方面的接触。

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.