Research and Teaching InterestsOur research is focused on elucidating the structure and function of proteins, with particular emphasis on fluorescent proteins and remodeling enzymes. The ultimate goal is to introduce novel biochemical, optical, or functional features into a protein’s scaffold. We utilize macromolecular X-ray crystallography to determine atomic-resolution structures, steady-state spectroscopy to investigate protein maturation kinetics, ultra-fast spectroscopy to examine the photochemistry of intrinsic chromophores, and molecular biology for protein engineering purposes.  Protein self-processing reactions: In fluorescent proteins such as GFP, the biosynthesis of several chemically distinct chromophores involves self-catalysis of peptide cyclization and oxidation reactions. Our research aims to develop step-by-step mechanistic models for the amino acid derivatizations that occur spontaneously upon protein folding, yielding brightly colored fluorophores from a single gene product. Structural basis of color evolution in fluorescent proteins: Four color classes (cyan, green, yellow, red) have evolved repeatedly and independently along different lineages of GFP-like proteins. We aim to understand how conformational remodeling facilitates phenotypic changes such as color switching events during protein evolution. Bioenergy and photosynthesis: The regulation of carbon fixation in higher plants and green algae includes remodeling enzymes responsible for maintaining the activity of RuBisCo, which catalyzes the central carboxylation reaction. We are interested in determining the mechanism of action of these auxiliary factors, as they appear to limit the net rate of CO2 fixation at elevated temperatures. Biomedical: We are investigating the structural and biochemical properties of M. tuberculosis protein factors with proposed roles in the regulation of MTB virulence. |
Publications
"An Improved Cerulean Flourescent Protein with Enhanced Brightness and Reduced Reversible Photoswitching," M. L. Markwardt, G.-J. Kremers, C. A. Kraft, K. Ray, P. J. C. Cranfill, K. A. Wilson, R. N. Day, R. M. Wachter, M. W. Davidson, M. A. Rizzo, PlosOne e17896 (2011)
"Atomic Resolution X-ray Structure of the Substrate Recognition Domain of Higher Plant Rubisco Activase," J. N. Henderson, A. M. Kuriata, R. Fromme, M. E. Salvucci, R. M. Wachter, J. Biol. Chem. Report in press (2011)
"Structural Changes Associated with the Acute Thermal Instability of Rubisco Activase," Barta, C., Dunkle, A. M., Wachter, R. M., Salvucci, M. E. , Archives of Biochemistry and Biophysics 499 17-25 (2010)
"Mechanistic Diversity of Red Fluorescence Acquisition by GFP-like Proteins (Current Topics Invited Review)," Wachter, R. M., Watkins, J. L., Kim, H., Biochemistry 49 7417-7427 (2010)
"Crystal Structure and Raman Studies of dsFP483, a Cyan Fluorescent Protein from Discosoma striata," Malo, G. D., Wang, M., Wu, D., Stellin, A., Tonge, P. J., Wachter, R. M., J. Mol. Biol. 378 869-884 (2008)
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