Department of Chemistry Biochemistry

REBEKKA WACHTER

Associate Professor
Ph.D., University of Oregon, 1996

Alexander von Humboldt Award for Experienced Researchers
NIH National Research Service Award

  Office: PSC-307  Lab: D-302, D-314
  Phone: (480)965-8188  Lab Phone: (480)727-7843
  Fax: (480) 965-2747
  Email: rwachter@asu.edu

 Dr. REBEKKA WACHTER's Lab or Group Website

Research and Teaching Interests

Our 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.

Selected Publications

"Purification and Biophysical Characterization of the CapA Membrane Protein FTT0807 from Francisella tularensis," Martin-Garcia, J., Hansen, D., Zook, J., Loskutov, A., Robida, M., Craciunescu, F, Sykes, K., Wachter, R. M., Fromme, P., Allen, J. P., Biochemistry (in press) (2014)

"Biophysical Characterization of Higher Plant Rubisco Activase," Henderson, J. N., Hazra, S., Dunkle, A. M., Salvucci, M. E., Wachter*, R. M., Biophys. Biochim. Acta 1834 87-97 (2013)

"The 1.6 A structure of a FRET-optimized Cerulean Fluorescent Protein," Watkins, J. L., Kim, H., Markwardt, M. L., Chen, L., Fromme, R., Rizzo, M. A., Wachter*, R. M., Acta Crystallogr. D 69 767-773 (2013)

"Ultrafast Proton Shuttling in Psammocora Cyan Fluorescent Protein," Kennis, J. T. M., van Stokkum, I. H. M., Peterson, D. S., Pandit, A., Wachter, R. M., The Journal of Physical Chemistry B 117 11134-11143 (2013)

"Activation of Interspecies-hybrid Rubisco Enzymes to Assess Different Models for the Rubisco Rubisco Activase Interaction ," Wachter, R. M., Salvucci, M. E., Carmo-Silva, A. E., Barta, C., Genkhov, T., Spreitzer, R. J., Photosynthesis Research 117 557-566 (2013)

"Acid-Base Catalysis and Crystal Structures of a Least-Evolved Ancestral GFP-like Protein Undergoing Green-to-Red Photoconversion ," Kim, H., Grunkemeyer, T. J., Modi, C., Chen, L., Fromme, R., Matz, M. V., Wachter*, R. M., Biochemistry 52 8048-8059 (2013)

"Protein Oligomerization Monitored by Fluorescence Fluctuation Spectroscopy: Self-Assembly of Rubisco Activase," Chakraborty, M., Kuriata, A. M., Henderson, J. N., Salvucci, M. E., Wachter*, R. M., Levitus*, M., Biophys. J. 103 949-958 (2012)

"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. 286 35683-35688 (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)

"Kinetic Isotope Effect Studies on the de novo Rate of Chromophore Formation in Fast- and Slow-maturing GFP Variants," Pouwels, L. J., Zhang, L., Chan, N. H., Dorrestein, P. C., Wachter, R. M., Biochemistry in press (2008)

"X-ray Structure of Cerulean GFP: A Tryptophan-Based Chromophore Useful for Fluorescence Lifetime Imaging," Malo, G. D., Pouwels, L. J., Wang, M., Weichsel, A., Montfort, W. R., Rizzo, M. A., Piston, D. W., Wachter, R. M., Biochemistry (Accelerated Publication) 46 9865-9873 (2007)

"Chromogenic Cross-link Formation in Green Fluorescent Protein ," Wachter, R. M., Acc. Chem. Res. 40 120-127 (2007)

"Reaction Progress of Chromophore Biogenesis in Green Fluorescent Protein," Zhang, L., Patel, H. N., Lappe, J. W., Wachter, R. M., J. Am. Chem. Soc. 128 4766-4772 (2006)

"The family of GFP-like proteins: Structure, Function, Photophysics and Biosensor Applications," Wachter, R. M., Photochem. Photobiol. 82 339-344 (2006)

"Development and Characterization of Green Fluorescent Protein Mutants with Altered Lifetimes," Scruggs, A. W., Flores, C. L., Wachter, R. M., Woodbury, N. W. , Biochemistry 44 13377- 13384 (2005)

"Maturation Efficiency, Trypsin Sensitivity and Optical Properties of Arg96, Glu222 and Gly67 Variants of Green Fluorescent Protein.," J.A. Sniegowski, M.E. Phail and R.M. Wachter, Biochem. Biophys. Res. Comm 332 657-663 (2005)

"Oxidative Chemistry in the GFP Active Site Leads to Covalent Cross-Linking of a Modified Leucine Side Chain with a Histidine Imidazole: Implications for the Mechanism of Chromophore Formation," M.A. Rosenow, H.N. Patel and R.M. Wachter, Biochemistry 44 8303-8311 (2005)

"Base-catalysis of Chromophore Fromation in Arg96 and Glu222 variants of Green Fluorescent Protein," J.A. Sniegowski, J.W. Lappe, H.N. Patel, H.A. Huffman and R.M. Wachter, J. Biol. Chem 280 26248-26255 (2005)

"zFP538, a Yellow Fluorescent Protein from Zoanthus, Contains a Novel Three-Ring Chromophore," S.J. Remington, R.M. Wachter, D.K. Yarbrough, B.P. Branchaud, D.C. Anderson, K. Kallio and K.A. Lukyanov, Biochemistry 44 202-212 (2005)

"X-ray diffraction analysis and molecular replacement solution of the cyan fluorescent protein dsFP483," M. Wang, H.N. Patel and R. Wachter, Acta Cryst. 61 922-924 (2005)

"The Crystal Structure of the Y66L Variant of Green Fluorescent Protein Supports a Cyclization-Oxidation-Dehydration Mechanism for Chromophore Maturation.," M.A. Rosenow, H.A. Huffman, M.E. Phail and R.M. Wachter, Biochemistry 43 4464-4472 (2004)

"Light Driven Decarboxylation of Wild-Type Green Fluorescent Protein.," Bell, A. F., Stoner-Ma, D., Wachter, R. M., Tonge, P. J. , J. Am. Chem. Soc. 125 6919-6926 (2003)