Wayne State University

Aim Higher



College of Liberal Arts & Sciences
Department of Chemistry
Faculty Page
 
Mary Kay Pflum
Title Associate Professor
Division Organic (Biochemistry)
Education B.A. Carleton College, 1992
Ph.D. Yale University, 1999
NIH Postdoctoral Fellow, Harvard University, 1999-2001
Office Chem 323
Phone (313)577-1515
E-Mail
Group http://chem.wayne.edu/pflumgroup


Research in the Pflum group integrates organic synthesis, biochemistry, and cell biology to understand the molecular basis of disease. For many years, we have been particularly interested in two proteins associated with cancer formation- kinase and histone deacetylase enzymes. Our goal is to use a chemical approach to characterize the role of these proteins in cancers, which will lead to identification of novel drugs targeting these proteins and biomarkers to diagnose illness.

Our group focuses on two main projects:

I. Kinase Enzymes

Kinases catalyze protein phosphorylation, which is a ubiquitous post-translational modification that can profoundly influence protein function. However, identifying the position of phosphorylated amino acids and the physiological significance of phosphorylation has been challenging due to the paucity of available tools. We are interested in developing chemical methods to characterize kinase-catalyzed post-translational modifications.

We recently reported use of ATP analogs to label, visualize, and enrich phosphorylated proteins from protein or peptide mixtures. The method relies upon ATP analogs with a gamma-phosphate tag that can be used as a cosubstrate by kinases to label recombinant proteins and cell lysates (Figure 1). Using this strategy, we have established kinase-catalyzed biotinylation, dansylation, and crosslinking (see Green and Pflum, 2007; Green and Pflum, 2009; Suwal and Pflum, 2010, Senevirathne and Pflum, 2013, Garre, et al. 2014). We are currently focused on applying these novel labeling methods to characterize phosphoproteins in cell lysates and live cells (see Fouda and Pflum, 2015). These studies will pioneer advances in cell signaling research by coupling cell biology and biochemistry with novel synthetic ATP analogs.

II. Histone Deacetylase Enzymes
Histone Deacetylase (HDAC) proteins are transcription factors that influence cell proliferation, differentiation, and cancer formation. In fact, several small molecule inhibitors of HDAC proteins are in clinical trials to treat cancer. Unfortunately, the known drugs typically interact with eleven human HDAC proteins nonspecifically. To overcome the limitation of available drugs, we use mutagenesis to characterize the activities of histone deacetylase 1 (HDAC1) (Figure 2A, see Weerasinghe, et al. 2008). We also have a medicinal chemistry program aimed at identifying isoform selective inhibitors (Figure 2B, see Choi et al. 2011; Weerasinghe et al. 2010; Bieliauskas et al. 2007). Our goal is to expand the utility of small molecule-based HDAC inhibitors in cancer research and treatment by combining synthetic organic chemistry and cell biology.






REPRESENTATIVE PUBLICATIONS

Ahmed E. Fouda and Mary Kay H. Pflum, A Cell Permeable ATP analog for Kinase-Catalyzed Biotinylation, Angewandte Chemie International Edition, 2015, in press.
http://dx.doi.org/10.1002/anie.201503041

Satish Garre, Chamara Senevirathne and Mary Kay H. Pflum, A comparative study of ATP analogs for phosphorylation-dependent kinase-substrate crosslinking, Bioorganic & Medicinal Chemistry, 2014, 1620-1625.
http://dx.doi.org/10.1016/j.bmc.2014.01.034

Magdalene K. Wambua, Dhanusha A. Nalawansha, Ahmed T. Negmeldin and Mary Kay H. Pflum, Mutagenesis Studies of the 14 Å Internal Cavity of Histone Deacetylase 1: Insights toward the Acetate-Escape Hypothesis and Selective Inhibitor Design, J. Med. Chem., 2014, 1620-1625.
http://pubs.acs.org/doi/full/10.1021/jm401837e

Chamara Senevirathne and Mary Kay H. Pflum, Biotinylated Phosphoproteins from Kinase-Catalyzed Biotinylation are Stable to Phosphatases: Implications for Phosphoproteomics, ChemBioChem, 2013, 381-387.
http://dx.doi.org/10.1016/j.bmcl.2012.09.093

Sun Ea Choi and Mary Kay H. Pflum, The structural requirements of histone deacetylase inhibitors: Suberoylanilide hydroxamic acid analogs modified at the C6 position, Bioorganic & Medicinal Chemistry Letters, 2012, 22, 7084-7086.
http://dx.doi.org/10.1016/j.bmcl.2012.09.093

Sun Ea Choi, Sujith V. W. Weerasinghe, and Mary Kay H. Pflum, The Structural Requirements of Histone Deacetylase Inhibitors: Suberoylanilide Hydroxamic Acid Analogues Modified at the C3 Position Display Isoform Selectivity, Bioorg Med Chem Lett, 2011, 21, 6139-6142.
http://dx.doi.org/10.1016/j.bmcl.2011.08.027

Sujit Suwal and Mary Kay H. Pflum, Phosphorylation-Dependent Kinase-Substrate Cross-Linking, Angewandte Chemie International Edition, 2010, 49(9), 1627-1630.
http://dx.doi.org/10.1002/anie.200905244

Keith D. Green and Mary Kay H. Pflum, Exploring Kinase Cosubstrate Promiscuity: Monitoring Kinase Activity through Dansylation, ChemBioChem, 2009, 10(2), 234-237.
http://dx.doi.org/10.1002/cbic.200800393

Sujith V. W. Weerasinghe, Guillermina Estiu, Olaf Wiest, and Mary Kay H. Pflum, Residues in the 11 Å Channel of Histone Deacetylase 1 Promote Catalytic Activity: Implications for Designing Isoform-Selective Histone Deacetylase Inhibitors, J. Med. Chem., 2008, 51(18), 5542-5551.
http://dx.doi.org/10.1021/jm800081j

Keith D. Green and Mary Kay H. Pflum, Kinase-Catalyzed Biotinylation for Phophoprotein Detection, J. Am. Chem. Soc., 2007, 129(1), 10-11.
http://dx.doi.org/10.1021/ja066828o

DEPARTMENT OF CHEMISTRY ©
5101 Cass Ave, Detroit, MI 48202
Phone: (313) 577-7784    Fax: (313) 577-8822