An award is made to support the acquisition of an automated Total Internal Reflection Fluorescence (TIRF) Microscope system to support the research, educational and outreach efforts at Marquette University (MU), Medical College of Wisconsin (MCW) and other research groups in the greater Milwaukee region. TIRF microscopy is an essential tool used to study, with high resolution, the mechanism and dynamics of macromolecular protein complex assembly and movement at cellular surfaces. TIRF has also been extensively adapted to study enzymes at single molecule resolution using various fluorescence-based approaches. The availability of a TIRF microscope at MU and neighboring institutions in the greater Milwaukee areas is required to provide hands-on training to undergraduate, graduate and postdoctoral researchers. The PI and co-PIs will provide training to all interested researchers, including students ranging in expertise from graduate to young undergraduate students. The PI and co-PIs have a strong record of training undergraduate and graduate students and the TIRF microscope will become a key part of the educational efforts at Marquette University and surrounding institutions in the greater Milwaukee region.
The projects underway in the laboratories of the PI, Co-PIs and other users fall under two broad categories of TIRF microscopy applications. The first group will use single molecule FRET to visualize the motions of fluorescently labelled enzymes and this information will be deconvoluted to understand their mechanism of action. The second group will utilize the TIRF microscope to study in vivo cellular processes, where the dynamics of fluorescently labeled proteins are followed within the cell. The TIRF microscope will be invaluable to: (1) performing mechanistic studies of ATP-dependent motor proteins; (2) clarifying the role of carrier domain movements in biotin-dependent metabolic enzymes; (3) identifying the assembly mechanism of prion formation on actin networks; (4) exploring the role of temperature in gamete development; (5) understand the molecular and cellular mechanisms of eukaryotic flagella; (6) studying Ras GTPase interactions on cellular membranes (7) describing the mechanism of G protein and β-arrestin chemokine receptor signaling; and (8) investigating the role of phosphorylation in β-arrestin and E3 ubiquitin ligase recruitment. Results from these studies will be published in peer-reviewed scientific journals, presented at scientific meetings, and used in both training and outreach activities.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
