This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).
Funds from the NSF MRI-R2 program were awarded to the University of Mississippi Medical Center to support the purchase of an Aviv AU-FDS Fluorescence Detection System. The new system is used to perform hydrodynamic, thermodynamic and macromolecular interaction studies on a wide range of cutting-edge biophysical projects by six major users. Projects include investigations of the interaction of proteins that regulate microtubule assembly, the aggregation of biologically important peptides, self association properties of kinesin molecular motor domains, self association of Hfq and its binding to sRNA and mRNA targets, aggregation properties of thermally responsive elastin-like polypeptide (ELP), and assembly properties of coilin, including its cellular size distribution. These projects will also result in the development of advanced Analytical Ultracentrifuge (AUC) methods and software. The Aviv Fluorescence Detection System brings capabilities to the AUC Facility that greatly enhance the kinds of studies and applications utilized by researchers. The projects enabled by the new equipment also greatly benefit and impact the students and post doctoral fellows involved in those projects by exposing them to state-of-the-art biophysical techniques. Investigators at UMMC are involved in several programs designed to encourage outreach and diversity in science. These include MARC (Minority Access to Research Careers), MFGN (Mississippi Functional Genomics Network), and the Base Pair Program for high school students in Jackson. In addition, undergraduates participate in summer research at UMMC supported by grant funds or SURE monies. The impact of the new equipment on projects at UMMC and throughout the state is further enhanced by the presence of the Mississippi Biophysical Consortium (MBC). Results of the research projects enabled by the AU-FDS system will be disseminated in peer-reviewed scientific journals, and by student and faculty presentations at regional and national meetings.
The goal of this research is to expand and develop the application of fluorescence detection optics (Aviv FDS) for Analytical Ultracentrifugation (AUC). AUC is a fundamental method that exposes macromolecules to a high rotational force and allows their sedimentation response to be recorded by various optical systems. The method allows the user to determine information about macromolecule purity, size, shape and association properties. This information is essential for understanding the basic behavior of macromolecules including proteins, nucleic acids (DNA and RNA), sugars, lipids, nanoparticles and a wide range of block co-polymers. The applications explored in this study include: characterization of a protein (ELP) drug delivery system for the treatment of cancer; the aggregation of β-amyloid to understand the mechanisms involved in Alzheimer’s; the association of an RNA binding protein called Hfq; determination of the size distribution of a cellular protein coilin as a function of phosphorylation; the size distribution of tubulin and microtubule polymers in cells in response to anticancer drug; the binding of DNA and siRNA to block-copolymers to development a nanoparticle based system for cancer treatment; the development of new drugs that bind to and inhibit specific DNA sequences that control mutated genes that cause cancer; dimerization of a microtubule binding protein EB1. The advantage of fluorescence detection is that it is highly selective for a specific probe or molecule even in a complex solution like cell extracts or human serum. Thus, the ELP drug delivery studies are focused on making measurements in serum to understand how these molecules behave in the actual treatment medium, human blood and serum. The coilin project investigates the size distribution in a cell extract. The tubulin project investigates the mechanism of action of anticancer drugs in cells as a means to understand both what is effective as a drug treatment but also why drug therapy might fail and how tumors become resistant to drugs. A major component of this project is the development of software tools that allow the analysis of FDS data and the theoretical understanding to model and interpret data collected in serum. In addition, this project has expanded the capabilities of the UMC AUC Facility and made FDS AUC a powerful resource for investigators throughout the state of Mississippi and collaborators throughout the US.
