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.
- Riegert, A. S., Narindoshvili, T., Coricello, A., Richards, N., & Raushel, F. M. (2022). Correction to “Functional Characterization of Two PLP-Dependent Enzymes Involved in Capsular Polysaccharide Biosynthesis from Campylobacter jejuni”. Biochemistry. 61(1), 46-46.
- Riegert, A. S., Narindoshvili, T., & Raushel, F. M. (2022). Discovery and Functional Characterization of a Clandestine ATP-Dependent Amidoligase in the Biosynthesis of the Capsular Polysaccharide from Campylobacter jejuni. Biochemistry. 61(2), 117-124.
- Riegert, A. S., Narindoshvili, T., Coricello, A., Richards, N., & Raushel, F. M. (2021). Functional Characterization of Two PLP-Dependent Enzymes Involved in Capsular Polysaccharide Biosynthesis from Campylobacter jejuni. Biochemistry. 60(37), 2836-2843.
- Bigley, A. N., Harvey, S. P., Narindoshvili, T., & Raushel, F. M. (2021). Substrate Analogues for the Enzyme-Catalyzed Detoxification of the Organophosphate Nerve Agents—Sarin, Soman, and Cyclosarin. Biochemistry. 60(38), 2875-2887.
- Holden, H. M., & Raushel, F. M. (2021). From the Three-Dimensional Structure of Phosphotriesterase.. Biochemistry. 60(46), 3413-3415.
- 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. Biophysical Journal. 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. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY. 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. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY. 242,
- Goble, A. M., Zhang, Z., Swaminathan, S., & Raushel, F. M. (2011). Identification and structure determination of novel substituted purine deaminase enzymes. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY. 242,
- Lund, L., Williams, H. J., & Raushel, F. M. (2010). Evidence for the utilization of formate as a substrate by carbamoyl phosphate synthetase. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY. 240,
- Raushel, F. M., & Bigley, A. N. (2019). Variants of phosphotriesterase for the hydrolysis and detoxification of nerve agents.
- Raushel, F. M., & Bigley, A. W. (2019). Variants of phosphotriesterase for the hydrolysis and detoxification of nerve agents.
- 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)