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

Hope College, Holland MI

Investigators

Abstract

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.