Enzymes catalyze a remarkable variety of chemical reactions with extremely high rate enhancements and very selective substrate specificity. The research efforts in our laboratory are directed towards a more complete understanding of the fundamental principles involved in enzyme-catalyzed chemistry and the dependence on protein structure. The pursuit of this information will provide the framework for the rational and combinatorial redesign of these complex molecules in an effort to exploit and develop the properties of enzyme active sites for a variety of chemical, biological, and medicinal uses. The techniques that we are using to solve these problems include steady-state and stopped-flow kinetics, NMR and EPR spectroscopy, X-ray crystallography, and the synthesis of inhibitors and suicide substrates. We are also using recombinant DNA methods to construct new proteins with novel catalytic properties. These efforts are currently being directed to the reactions catalyzed by phosphotriesterase and enzymes involves in the degradation of lignin and the metabolism of novel carbohydrates from the human gut microbiome.
The phosphotriesterase enzyme catalyzes the hydrolysis of organophosphate insecticides and other toxic organophosphate nerve agents. We have discovered that the active site of this protein consists of a unique binuclear metal center for the activation of water. We are now investigating the structure and properties of this metal center as a model system for the evolution of enzyme structure and function. Toward this end we have mutated the active site of this enzyme in a research project to create novel enzymes with the ability to detect, destroy, and detoxify various chemical warfare agents such as sarin, soman, and VX. The Raushel laboratory is also engaged in a large scale research project that is focused on the development of novel strategies for the discovery of new enzymes.
- Mabanglo, M. F., Huddleston, J. P., Mukherjee, K., Taylor, Z. W., & Raushel, F. M. (2020). Structure and Reaction Mechanism of YcjR, an Epimerase That Facilitates the Interconversion of d-Gulosides to d-Glucosides in Escherichia coli.. BIOCHEMISTRY. 59(22), 2069-2077.
- Huddleston, J. P., & Raushel, F. M. (2020). Functional Characterization of Cj1427, a Unique Ping-Pong Dehydrogenase Responsible for the Oxidation of GDP-d-glycero-α-d-manno-heptose in Campylobacter jejuni.. BIOCHEMISTRY.
- Bigley, A. N., Narindoshvili, T., Xiang, D. F., & Raushel, F. M. (2020). Stereoselective Formation of Multiple Reaction Products by the Phosphotriesterase from Sphingobium sp. TCM1.. BIOCHEMISTRY. 59(12), 1273-1288.
- Huddleston, J. P., Anderson, T. K., Spencer, K. D., Thoden, J. B., Raushel, F. M., & Holden, H. M. (2020). Structural Analysis of Cj1427, an Essential NAD-Dependent Dehydrogenase for the Biosynthesis of the Heptose Residues in the Capsular Polysaccharides of Campylobacter jejuni.. BIOCHEMISTRY.
- Huddleston, J. P., & Raushel, F. M. (2019). Biosynthesis of GDP-D-glycero-alpha-D-manno-heptose for the Capsular Polysaccharide of Campylobacter jejuni. BIOCHEMISTRY. 58(37), 3893-3902.
- Yang, K., Ren, Z., Raushel, F. M., & Zhang, J. (2016). Structures of the Carbon-Phosphorus Lyase Complex Reveal the Binding Mode of the NBD-Like PhnK. Biophys J. 110(3), 159A-159A.
- Hobbs, M. E., Williams, H. J., & Raushel, F. M. (2012). Discovery of an L-fucono-1,5-lactonase within cog3618 from the amidohydrolase superfamily. ACS Photonics. 244,
- Ornelas, A., Narindoshvili, T., Sugadev, R., Kumaran, D., Swaminathan, S., & Raushel, F. M. (2011). Characterization of members of the amidohydrolase superfamily in COG3964: Determining function of Atu3266 and Ef0837. ACS Photonics. 242,
- Goble, A. M., Zhang, Z., Swaminathan, S., & Raushel, F. M. (2011). Identification and structure determination of novel substituted purine deaminase enzymes. ACS Photonics. 242,
- Lund, L., Williams, H. J., & Raushel, F. M. (2010). Evidence for the utilization of formate as a substrate by carbamoyl phosphate synthetase. ACS Photonics. 240,
- Ren, Zhongjie (2016-12). Structural and Mechanistic Characterization of Carbon-Phosphorus Lyase Multi-Protein Complex and the Phosphotriesterase from Sphingobium sp. Strain TCM1. (Doctoral Dissertation)
- Ghodge, Swapnil Vijay (2015-05). Mechanistic Characterization and Function Discovery of Phosphohydrolase Enzymes from the Amidohydrolase Superfamily. (Doctoral Dissertation)
- Vladimirova, Anna V (2015-05). Enzymes in COG2159 of the Amidohydrolase Superfamily: Structure and Mechanism of 5-Carboxyvanillate Decarboxylase (LIGW). (Doctoral Dissertation)
- Ragavan, Mukundan (2014-08). Dissolution Dynamic Nuclear Polarization of Polypeptides. (Doctoral Dissertation)
- Hobbs, Merlin Eric (2014-05). Mechanistic and Functional Characterization of Lactonases of COG3618 in the Amidohydrolase Superfamily. (Doctoral Dissertation)