Areas of Interest
Our overall research goal is to understand the mechanisms used by biological catalysts, both proteins and nucleic acids, to achieve high efficiency and stringent specificity. We are particularly focused on the mechanism of medically important metalloenzymes. We are investigating the catalytic mechanism and specificity of protein farnesyltransferase and protein geranylgeranyltransferase I. These enzymes catalyze the addition of a prenyl group onto a C-terminal cysteine on a variety of substrates involved in signal transduction. Compounds that inhibit FTase are being investigated as possible antitumor agents. A second enzyme, UDP-3-O-acyl-GlcNAC deacetylase (LpxC) is a zinc metalloenzyme that catalyzes the first committed step in the pathway to form Lipid A, a crucial component of the outer membrane of gram negative bacteria. Inhibitors of this enzyme have antibacterial activity. To further the development of novel inhibitors we are elucidating detailed structure-function relationships in the active site of these proteins using mutagenesis, kinetic analysis, X-ray crystallography, and spectroscopic studies. Additionally, we have begun mechanistic studies of two related enzymes, histone deacetylase and protein palmitoyltransferase. For all of the metalloenzymes, a key question is the identify of the in vivo metal ion and whether metal switching is an important regulatory mechanism. Finally, we are developing methods for high-throughput screens of protein-bound transition metal ions for use in assaying all of the proteins in the yeast proteome to identify the yeast "metallome".
We also investigating the catalytic modes of ribozymes compared to proteins by determining the structure and mechanism of ribonuclease P (RNase P), a ribonucleoprotein complex that catalyzes the cleavage of tRNA precursors. We have demonstrated that the protein component enhances the catalytic efficiency by interacting with both P RNA and pre-tRNA. In the future, we will elucidate the structure of the holoenzyme using fluorescence resonance energy transfer, crystallography and spectroscopy. Finally, we will investigate the mechanism of yeast RNase P (in collaboration with Dr. D. Engelke) which contains one RNA and multiple protein subunits and purify, clone and characterize the RNase P from mammalian mitochondria which is proposed to be a protein catalyst.
Zinc, iron and copper ions are proposed to play important biological roles, especially in neurobiology, as well as playing important roles in the development of a number of diseases, including diabetes. Furthermore, a number of metals, such as lead and cadmium, are toxic. We are investigating the mechanisms of metal homeostasis and metal toxicity using a combination of biochemistry, genetics and imaging. To this end, we are redesigning the zinc metalloenzyme, carbonic anhydrase II, to optimize a fluorescent biosensor for measuring and imaging "readily exchangeable" metal ions in complex biological mixtures, such as cells, plasma and sea water and are developing similar sensors to measure cellular iron concentrations. Additionally, we are using X-ray fluorescence microprobe imaging to image total metal ions in wild-type and mutant yeast cells. These imaging methods are being used to understand basic mechanisms of metal homeostasis. Finally, we are examining the metal content and the mechanisms of metal insertion into proteins in vivo using biochemistry and analysis of libraries of deletion mutants. These studies should lead to a better understanding of the functions and regulation of biological transition metals.
Honors & Awards
Mildred Cohn Award in Biological Chemistry, ASBMB, 2020
Gordon Hammes Distinguished Lectureship, American Chemical Society, 2016
Award for Encouraging Women into Careers in the Chemical Sciences, American Chemical Society, 2016
Emil Thomas Kaiser Award, Protein Society, 2014
Distinguished University Professorship, University of Michigan 2013
Repligen Award, American Chemical Society, 2012
Elected Fellow, American Association for the Advancement of Science, 2007
Distinguished Faculty Achievement Award, University of Michigan, 2005
Sarah Power Goddard Award, 2005
Jerome and Isabella Karle Collegiate Professor, University of Michigan, 2003
Faculty Recognition Award, University of Michigan, 2001
American Heart Association Established Investigator Award, 1992–1997
Phosphorylation of Histone Deacetylase 8: Structural and Mechanistic Analysis of the Phosphomimetic S39E Mutant.
Welker Leng KR, Castañeda CA, Decroos C, Islam B, Haider SM, Christianson DW, Fierke CA.
Biochemistry. 2019; 58: 4480–93.
Interplay between substrate recognition, 5' end tRNA processing and methylation activity of human mitochondrial RNase P.
Karasik A, Fierke CA, Koutmos M.
RNA. 2019; 25: 1646–60.
The chaperone SmgGDS-607 has a dual role, both activating and inhibiting farnesylation of small GTPases.
García-Torres D, Fierke CA.
J Biol Chem. 2019; 294: 11793–804.
A cationic polymethacrylate-copolymer acts as an agonist for β-amyloid and an antagonist for amylin fibrillation.
Sahoo BR, Genjo T, Nakayama TW, Stoddard AK, Ando T, Yasuhara K, Fierke CA, Ramamoorthy A.
Chem Sci. 2019; 10: 3976–86.
Alzheimer's amyloid-beta intermediates generated using polymer-nanodiscs.
Sahoo BR, Genjo T, Bekier M, Cox SJ, Stoddard AK, Ivanova M, Yasuhara K, Fierke CA, Wang Y, Ramamoorthy A.
Chem Commun. 2018; 54: 12883–86.
Conservation of coactivator engagement mechanism enables small-molecule allosteric modulators.
Henderson AR, Henley MJ, Foster NJ, Peiffer AL, Beyersdorf MS, Stanford KD, Sturlis SM, Linhares BM, Hill ZB, Wells JA, Cierpicki T, Brooks CL 3rd, Fierke CA, Mapp AK.
Proc Natl Acad Sci USA. 2018; 115: 8960–65.
SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent.
Jennings BC, Lawton AJ, Rizk Z, Fierke CA.
Biochemistry. 2018; 57: 4289–98.
For a list of publications from PubMed, click HERE