Carol Fierke

Our goal is to understand the mechanisms used by biological catalysts, both proteins and nucleic acids, to achieve high efficiency, stringent specificity and rigorous control. An understanding of these principles is essential for: understanding biological catalysis in vivo, designing novel inhibitors for therapeutic use, and developing novel catalysts for a variety of tasks, including organic synthesis and quantitative analysis of complex mixtures. We are elucidating catalytic mechanisms and essential active site features of metalloenzymes and ribozymes, including protein farnesyltransferase, UDP-3-O-acyl-GlcNAC deacetylase, histone deacetylase and ribonuclease P. These studies should enhance our ability to design potent inhibitors of these enzymes useful for the treatment of cancer or bacterial infections. In particular, we are investigating the role of proteins in modulating the reactivity of bound Zn(II) and developing methods for identifying novel metal sites in proteins. We are also investigating the biological importance of protein prenylation and acetylation. Finally, we are elucidating the role of metal ions and protein/RNA interactions in ribonuclease P, a ribozyme/protein complex. These studies are increasing our understanding of the catalytic modes used by ribozymes in comparison to protein catalysts.

We are testing our understanding of biological catalysis by the rational design or redesign of an enzyme. To this end, we are redesigning the zinc metalloenzyme, carbonic anhydrase II, to optimize a fluorescent biosensor for measuring and imaging metal ions in complex biological mixtures. Zinc ions are proposed to play important signaling roles in vivo, especially in neurobiology, which can investigated using novel imaging methods. Additionally, we are using "directed evolution" approaches to prepare and identify aldolase variants with novel substrate specificities. These aldolase variants will be used to improve the utility of these enzymes as biocatalysts for organic synthetic reactions. Characterization of the structure and function of these novel proteins will provide insights into catalysis, molecular recognition and molecular evolution.

Awards

Emil Thomas Kaiser Award, 2014
Jerome and Isabella Karle Distinguished University Professor of Chemistry, 2013
ACS Repligen Award in Chemistry of Biological Processes, 2011
Rackham Distinguished Graduate Mentoring Award, 2011
Distinguished Faculty Achievement Award, 2005
Power Award, 2005
American Cancer Society Junior Faculty Research Award    
American Heart Association Established Investigator Award    

Representative Publications

Bozym, R. A., Thompson, R. B., Stoddard, A. K. and Fierke, C. A. (2006) Measuring picomolar intracellular exchangeable zinc in PC-12 cells using a ratiometric fluorescence biosensor, ACS Chemical Biology 1, 103-111.

**Pais, J. E., Bowers, K. E. and Fierke, C. A. (2006) Measurement of the a-Secondary Kinetic Isotope Effect for the Reaction Catalyzed by Mammalian Protein Farnesyltransferase, J. Am. Chem. Soc. 128, 15086-7.

**Getz, M. M., Andrews, A. J., Fierke, C. A. and Al-Hashimi, H. M. (2007) Structural Plasticity and Mg2+ Binding Properties of RNase P P4 from Combined Analysis of NMR Residual Dipolar Couplings and Motionally Decoupled Spin Relaxation, RNA 13, 251-66.

**Niranjanakumari, S., Day-Storms, J. J., Ahmed, M., Hsieh, J., Zahler, N. H., Venters, R. A. and Fierke, C. A. (2007) Probing the Architecture of the B. subtilis RNase P Holoenzyme Active Site by Crosslinking and Affinity Cleavage, RNA 13, 521-535.

Lipton, A. S., Heck, R. W., Hernick, M., Fierke, C. A. and Ellis, P. D. (2008) Residue Ionization in LpxC Directly Observed by 67Zn NMR Spectroscopy, J. Am. Chem. Soc 130,12671-9. http://www.ncbi.nlm.nih.gov/pubmed/18761443

Hsieh, J., Walker, S.C., Fierke, C.A. and Engelke, D. R. (2009) Pre-tRNA Turnover Catalyzed by the Yeast Nuclear RNase P Holoenzyme is Limited by Product Release, RNA, in press. http://www.ncbi.nlm.nih.gov/pubmed/19095620

Dowling, D.P., Gantt, S.L., Gattis, S.G., Fierke, C.A., and Christianson, D.W. (2008) Structural studies of human histone deacetylase 8 and its site-specific variants complexed with substrate and inhibitors, Biochemistry 47, 13554-63. http://www.ncbi.nlm.nih.gov/pubmed/19053282

Hougland, J.L., Scott, S.A., Gibbs, R. A. and Fierke, C.A. (2009) Context-dependent substrate recognition by protein farnesyltransferase, Biochemistry, in press.

Research Areas(s)

• Analytical Chemistry
  Bioanalytical Chemistry
  Bioinorganic Chemistry
  Biophysical Chemistry
  Chemical Biology
  Nano Chemistry
  Optics and Imaging
  Organic Chemistry
  RNA BioChemistry
  Sensor Science

Other Research Interests

• Biological Catalysis
  Molecular Recognition
  Enzyme Engineering

Other Postions

• Enzymes, Coenzymes and Metabolic Pathways Gordon Conference Editorial Board, RNA, Chair, 2005
• Biological Chemistry Division, American Chemical Society, Chair