RUI: Quantifying Fluorescent Probe Dynamics in FRET Experiments by Combining Single-Molecule and Bulk Spectroscopies with Molecular Dynamics and Quantum Mechanics Simulations

在化学系化学测量和成像计划的支持下，霍普学院的Brent Kruger教授和他的团队试图提高人们对荧光检测共振能量转移(FRET)的理解和能力，FRET是一种表征蛋白质和核酸等生物重要分子的结构和动力学的强大方法。作为测试系统，基于发夹核酶的几个生物学上重要但相对简单的DNA和RNA序列将被一系列荧光探针标记，以考察探针分子和化学结合方法对FRET结果的影响。将使用荧光测量和复杂的计算建模来提供对结构动力学的详细了解。由于FRET是一种在化学、生物学和生物物理学中广泛使用的技术，这项工作的结果将对一大批研究人员产生积极影响。本研究将为参与研究的学生提供有效的研究性训练经验。通过霍普学院已有的项目，除了霍普学院的本科生外，这些学生还将包括两年制的大学和当地的高中生。在资助期内，每年夏季将有四至五名学生参与研究，其中至少包括一名代表不足的少数族裔学生。此外，一个外展计划每年夏天将有12名高中教师来到霍普的校园，学习分子建模技术，并为他们的教室开发符合州和国家教育标准的课程。对过去两年从这些课程中受益的学生的评估显示，40%的学生报告对上大学的兴趣增加，29%的学生报告对从事STEM职业的兴趣增加。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Brent Kruger and his group at Hope College seek to improve the understanding and therefore the capabilities of fluorescence-detected resonance energy transfer (FRET), a powerful method for characterizing the structure and dynamics of biologically important molecules like proteins and nucleic acids. As test systems, several biologically important yet relatively simple DNA and RNA sequences based on the hairpin ribozyme will be tagged with a series of fluorescent probes to examine the impact of probe molecule and chemical attachment method on FRET results. Both fluorescence measurements and sophisticated computational modeling will be used to provide detailed understanding of structural dynamics. Because FRET is a widely-used technique in chemistry, biology, and biophysics, the results of this work will positively impact a large community of researchers. The research will provide effective research training experiences for the students involved. Through programs already in place at Hope College, these will include two-year-college and local high school students in addition to Hope College undergraduates. Over the period of the grant, four to five students will be engaged in research each summer including at least one underrepresented minority student. In addition, an outreach program will bring 12 high school teachers to Hope's campus each summer to learn molecular modeling techniques and to develop lessons for their classrooms that meet state and national education standards. Assessment of the students who have benefitted from these lessons over the past two years shows that 40% of students report an increased interest in attending college and 29% report an increased interest in pursuing a STEM career.