Our laboratory is interested in the mechanisms by which enzymes are regulated in the cell. In particular, we are interested in allosteric regulation of enzyme activity. Consequently, we are interested in understanding the nature of the conformational change in proteins that can be effected by the binding of ligands, and specifically how these changes alter the catalytic behavior of enzymes subject to allosteric regulation. We endeavor to investigate properties that are complementary to those determined by x-ray crystallography in order to develop a comprehensive picture of the structure-function relationships involved in the regulatory phenomenon. For example, we are interested in how the dynamics of protein structure might dictate the nature of an allosteric effect. Techniques and approaches that we use in the laboratory include analysis of enzyme kinetics; analysis of the thermodynamics of enzyme-ligand interactions; time-resolved and steady-state fluorescence spectroscopy; analysis of the effects of temperature and hydrostatic pressure (up to 4 kbar) on enzyme properties, site-specific mutagenesis, isothermal titration calorimetry, and molecular graphics.
- Whitaker, A. M., & Reinhart, G. D. (2016). The effect of introducing small cavities on the allosteric inhibition of phosphofructokinase from Bacillus stearothermophilus. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS. 607, 1-6.
- McGresham, M. S., & Reinhart, G. D. (2015). Enhancing allosteric inhibition in Thermus thermophilus phosphofructokinase. BIOCHEMISTRY. 54(3), 952-958.
- McGresham, M. S., Lovingshimer, M., & Reinhart, G. D. (2014). Allosteric regulation in phosphofructokinase from the extreme thermophile thermus thermophilus. BIOCHEMISTRY. 53(1), 270-278.
- Ranjit, S., Dvornikov, A., Holland, D. A., Reinhart, G. D., Jameson, D. M., & Gratton, E. (2014). Application of three-photon excitation FCS to the study of protein oligomerization. The journal of physical chemistry. B. 118(50), 14627-14631.
- Mosser, R., Reddy, M., Bruning, J. B., Sacchettini, J. C., & Reinhart, G. D. (2013). Redefining the Role of the Quaternary Shift in Bacillus stearothermophilus Phosphofructokinase. BIOCHEMISTRY. 52(32), 5421-5429.
- Tindall, A. J., & Reinhart, G. D. (2016). Kinetic Characterization of Human Liver Phosphofructokinase. Biophys J. 110(3), 398A-398A.
- Holland, D. A., & Reinhart, G. D. (2016). Use of 2-Photon Fluorescence Correlation Spectroscopy to Investigate Rat Liver Phosphofructokinase Self-Association. Biophys J. 110(3), 217A-217A.
- Tian, X., Lasagna, M., & Reinhart, G. D. (2015). Identifying Unique Conformations in Thermus Thermophilus Phosphofructokinase using Fluorescence Phasors. Biophys J. 108(2), 212A-212A.
- Tindall, A., & Reinhart, G. D. (2015). Kinetic Characterization of Human Liver Phosphofructokinase. Biophys J. 108(2), 532A-532A.
- Whitaker, A. M., Naik, M. T., & Reinhart, G. D. (2015). Propogation of the Allosteric Signal in Bacillus Stearothermophilus Phosphofructokinase Examined by Methyl-TROSY NMR. Biophys J. 108(2), 30A-31A.
- Tian, Xinxin (2016-12). Probing the Details of the Allosteric Inhibition in Phosphofructokinase from Thermus thermophilus. (Doctoral Dissertation)
- Tindall, Amanda Jean (2016-05). Expression, Purification, and Kinetic Characterization of Human Liver Phosphofructokinase. (Master's Thesis)
- Whitaker, Amy Michelle (2015-12). Exploration of Thermodynamic and Structural Changes Relevant to the Allosteric Inhibition in Phosphofructokinase from Bacillus Stearothermophilus. (Doctoral Dissertation)
- Laird, Bobby Wayne (2015-05). Distinguishing Allosteric Interactions Using Thermodynamic Analysis and the First Reported Crystal Structure of Phospho(Enol)Pyruvate Bound E. (coli Phosphofructokinase. Doctoral Dissertation)
- Perez, Stephanie (2012-12). Illuminating the Heterotropic Communication of the Pair-wise Interactions in Phosphofructokinase from Bacillus stearothermophilus. (Doctoral Dissertation)