Chemical Biology & Spectroscopy
| Enzymology and Inhibition of Antibacterial Drug Targets | |
 |
Much of the work in the Tonge lab involves detailed studies into the mechanism of enzymes that are known or proposed drug targets for therapeutic intervention. We use spectroscopic techniques such as Raman, FTIR and NMR spectroscopy coupled with X-ray crystallography and site directed mutagenesis to obtain fundamental insight into how enzymes cause and stabilize charge rearrangement. A key aspect of this work is the design and synthesis of substrate analogues which are used to develop structure-reactivity relationships for enzyme inhibition. This information is used in turn to rationally design and synthesize high affinity enzyme inhibitors. Compounds are screened for antibacterial activity and cytotoxictity, and the intracellular mode of action os elected compounds is evaluated using DNA microarrays, target overexpression and photoaffinity labeling. Compounds that progress through the primary screens will be evaluated in vivo for toxicity, bioavailability, pharmacodynamics, pharmacokinetics and antibacterial activity in animal models of infection. |
| Mycobacterium Tuberculosis : Fatty acid Biosynthesis, Menaquinone Biosynthesis, Cell Division Francisella Tularensis and Staphylococcus Aureus: Fatty Acid Biosynthesis |
|
| Fluorescent Proteins and Light-Activated Proteins | |
Green fluorescent proteins (GFPs) are intrinsically fluorescent and have a wide range of applications in molecular and cell biology. Importantly, it has been shown that light absorption causes structural changes in the GFP chromophore that result in alterations in the chromophore’s fluorescence. We are using a variety of spectroscopic methods, including ultrafast vibrational spectroscopy, coupled with site directed mutagenesis to determine how the protein environment controls the spectroscopic properties of the chromophore. In addition, we are constructing an expression system to introduce isotopic labels and unnatural amino acids into the chromophore in order to study the mechanism of chromophore formation. These experiments will result in novel GFPs with faster rates of chromophore formation. |
|
 |
 |
GFP |
AppA: Light Driven H-Bond Rearrangement |
