Grazyna D. Szklarz, Ph.D. Associate Professor Department of Basic Pharmaceutical Sciences West Virginia University P.O. Box 9530 Morgantown, WV 26506-9530 Phone: (304) 293-1473 Fax: (304) 293-2576 E-Mail: gszklarz@hsc.wvu.edu Link to Full CV - PDF

Education:

Post-doctoral Training

Postdoctoral fellow, 1991-92, University of California, San Francisco, California

Graduate Training

M.S. in Biology, 1981, Maria Curie-Sklodowska University, Lublin, Poland
Ph.D. in Chemistry, 1991, Clarkson University, Potsdam, New York

Positions:

Post-doctoral Fellow, 1991-1992, Department of Pediatrics, University of California, San Francisco, CA

Associate Research Scientist, 1992-1995, Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ

Assistant Research Scientist, 1995-1997, Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ

Associate Research Scientist, 1998, Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ

Assistant Professor, August 1998-2004, Department of Basic Pharmaceutical Sciences, School of Pharmacy, West Virginia University, Morgantown, WV

Associate Professor, July 2004-present, Department of Basic Pharmaceutical Sciences, School of Pharmacy, West Virginia University, Morgantown, WV

Research Interests:

Structure and function of cytochromes P450
Homology modeling of enzyme structure
Utilization of molecular modeling methods to study protein function
P450-mediated drug metabolism and carcinogenesis

Research Summary

My current research focuses on structure-function relationships of human cytochromes P450 1A1 and 1A2, which play a key role in the metabolic activation of chemical carcinogens and in the metabolism of drugs. The objective of this research is to elucidate the structural basis for the function of these enzymes using a combination of molecular modeling and experimental methods, such as heterologous expression, site-directed mutagenesis and biochemical assays. We apply molecular modeling methods, such as molecular dynamics, to predict product profiles and kinetic constants, as well as NMR techniques, which will help to characterize enzyme-substrate interactions in the active site.

This research lies at the interface between the application of the theory of computational chemistry and the utilization of experimental methods, such as NMR and molecular biology techniques, and has a potential for advancing our understanding of how enzymes activate or deactivate drugs and xenobiotics.

Research Techniques:

Molecular modeling , homology modeling of proteins
Heterologous expression of mammalian enzymes in E. coli
Protein purification and analysis - chromatography, SDS-PAGE, Western blots, etc.
Enzyme assays, enzyme kinetics - spectroscopic methods, HPLC
Molecular biology - gene cloning and site-directed mutagenesis

Recent/Current Grants:

Structural basis for functional differences between human cytochromes P450 1A1 and 1A2, 07/01/99- 06/30/00 (WVU Senate Research Grant)

Active site model of cytochromes P450 1A1 and 1A2, 08/15/00-07/31/02 (NIH/NCI)
Structural determinants of P450 1A1 and 1A2 specificity. 09/14/01-08/31/06 (NIH/NCRR)

Cytochrome P450-reductase interactions in the P450 1A subfamily, 9/15/06-11/30/06 (Bridge Grant, WVU HSC)

Cytochrome P450-reductase interactions in P450 1A subfamily, 02/02/07-01/31/10 (NIH/NIGMS)

Recent Publications:

2008

Tu, Y., Deshmukh, R., Sivaneri, M. and Szklarz, G.D.  2008.  The application of molecular modeling for prediction of substrate specificity in cytochrome P450 1A2 mutants.  Drug Metab. Disp. 36: 2371-2380.

2005 and before

Ericksen, S.S. and Szklarz, G.D. 2005. Regiospecificity of human cytochrome P450 1A1-mediated oxidations: the role of steric effects. J. Biomol. Struct. Dyn., 23:243-256.

Schwarz, D., Kisselev, P., Ericksen, S.S., Szklarz, G.D., Honeck, H., Schunck, W.-H., and Roots, I. 2004. Arachidonic and eicosapentaenoic acid metabolism by human CYP1A1. Biochem. Pharmacol., 67:1445-1457.

Liu, J., Ericksen, S.S., Sivaneri, M., Besspiata, D., Fisher, C.W. and Szklarz, G.D. 2004. The effect of reciprocal active site mutations in human cytochromes P450 1A1 and 1A2 on alkoxyresorufin metabolism. Arch. Biochem. Biophys., 424:33-43.

Liu, J., Ericksen, S.S., Besspiata, D., Fisher, C.W. and Szklarz, G.D. 2003. Characterization of substrate binding to cytochrome P450 1A1 using molecular modeling and kinetic analyses: case of residue 382. Drug Metab. Disp. 31:412-420.

Szklarz, G.D. and Paulsen, M.D.  2002.  Molecular modeling of cytochrome P450 1A1: Enzyme-substrate interactions and substrate binding affinities.  J. Biomol. Struct. Dyn. 20: 155-162.

Xue, L., Wang, H.F., Wang, Q., Szklarz, G.D., Domanski, T.L., Halpert, J.R. and Correia, M.A. 2001.  Influence of P450 3A4 SRS-2 residues on cooperativity and/or regioselectivity of aflatoxin B1 oxidation.  Chem. Res. Toxicol. 14: 483-491.

Szklarz, G.D., Graham, S.E. and Paulsen, M.D. 2000.  Molecular modeling of mammalian cytochromes P450: application to study enzyme function. In: Vitamins and Hormones, G. Litwack, ed., Academic Press, vol. 58, pp. 53-86.

Strobel, S.M., Szklarz, G.D., He, Y., Foroozesh, M., Alworth, W.L., Roberts, E.S., Hollenberg P.F. and Halpert, J.R. 1999. Identification of selective mechanism-based inactivators of cytochromes P450 2B4 and 2B5, and determination of the molecular basis for differential susceptibility.  J. Pharmacol. Exp. Therap., 290: 445-451.

He, Y.Q., Harlow, G.R., Szklarz, G.D., and Halpert, J.R. 1998. Structural determinants of progesterone hydroxylation by cytochrome P450 2B5: the role of nonsubstrate recognition site residues.  Arch. Biochem.Biophys. 350:333-339.

Kobayashi, Y., Fang, X., Szklarz, G.D., and Halpert, J.R. 1998. Probing active site of cytochrome P450 2B1: metabolism of 7-alkoxycoumarins by the wild-type and five site-directed mutants.  Biochemistry 37: 6679-6688.

Wang, H., Dick, R., Yin H., Licad-Coles, E., Kroentz, D.L., Szklarz, G., Harlow, G., Halpert, J.R. and Correia, M.A. 1998. Structure-function relationships of human liver cytochromes P450 3A: aflatoxin B1 metabolism as a probe. Biochemistry 37: 12536-12545.

Szklarz, G.D. and Halpert, J.R. 1998. Molecular basis of P450 inhibition and activation: implications for drug development and drug therapy. Drug Metab. Disp., 26:1179-1184.

Teaching:

Professional:

Phar 731: Biopharmaceutics and Pharmacokinetics

Graduate:

Phar 779: Drugs Discovery and Development
Phar 780: Introduction to Molecular Modeling
Phar 781: Drug Metabolism

Special Service:

Member of Editorial Board of Drug Metabolism and Disposition
Workshop Presenter for Expanding Your Horizons in Math and Science Conferences organized by AWIS-WV

Professional Memberships:

American Association of Colleges of Pharmacy
American Chemical Society
American Society for Pharmacology and Therapeutics
Association for Women in Science
International Society for the Study of Xenobiotics