department of pharmacology

John Mieyal, Ph.D.

Mieyal

Professor
Department Vice Chair
Director of the MTTP

Phone: (216) 368-3383
Fax: 216-368-1300
E-mail: john.mieyal@case.edu
RT300 Research Tower

Research

Modulation of the thiol-disulfide status of critical cysteine residues on proteins is becoming recognized as an important mechanism of oxidative signal transduction as well as an important consequence of oxidative stress associated with aging and various disease states, including cardiovascular and neurodegenerative diseases, diabetes, AIDS, and cancer. Within these various contexts, a prevalent form of cysteine modification is reversible formation of protein mixed disulfides (protein-SSG) with intracellular glutathione (GSH).

Our laboratory is focused on the molecular mechanisms and physiological implications of enzymes that catalyze thiol-disulfide oxidoreductase (TDOR) reactions. In particular, we have characterized glutaredoxin (thioltransferase) as the TDOR enzyme that displays specificity and high catalytic efficiency for protein-SSG substrates, including hemoglobin-SSG, NF1-SSG, HIV-protease-SSG, and actin-SSG. This realization has placed made glutaredoxin a focal point in advancing understanding of protein-S-glutathionylation as a regulatory mechanism akin to phosphorylation of proteins. We are employing a range of cellular, molecular, and structural biology approaches to delineate the molecular basis for glutaredoxin catalysis and its role in regulation of fundamental cellular processes like proliferation, differentiation and apoptosis. A key objective in our research program is to characterize also the mechanisms of formation of specific protein-SSG intermediates and identify the enzymes responsible for catalyzing these reactions within cells. We are also focused on delineating changes in the regulation of protein-SSG status of specific effector proteins associated with the various disease states that involve oxidative stress.


PostDoctoral Positions Available


Selected References:

 

Gallogly MM, Shelton MD, Qanungo S, Pai HV, Starke DW, Hoppel CL, Lesnefsky EJ, and Mieyal JJ (2010), Glutaredoxin (Grx1) regulates apopotosis in cardiomyocytes via NFκB targets Bcl-2 and Bcl-xL: implications for cardiac aging, Antioxidants & Redox Signaling, [Epub Nov. 25, 2009], in press.

Shelton MD, Distler AM, Kern TS, Mieyal JJ. (2009), Glutaredoxin regulates autocrine and paracrine pro-inflammatory responses in retinal glial (Muller) cells, J Biol Chem. 284, 4760-4766.

Gallogly, M.M., D.W. Starke, and J.J. Mieyal, Mechanistic and Kinetic Details of Catalysis of Thiol-Disulfide Exchange by Glutaredoxins and Potential Mechanisms of Regulation, invited forum review article in Antioxidants & Redox Signaling, 11, 1059-1081 (2009).

Zhu X, Gallogly MM, Mieyal JJ, Anderson VE, Sayre LM, Covalent cross-linking of glutathione and carnosine to proteins by 4-oxo-2-nonenal, Chem. Res. Toxicol. 22,1050-1059 (2009).

Park JW, Mieyal JJ, Rhee SG, Chock PB. (2009) Deglutathionylation of 2-Cys peroxiredoxin is specifically catalyzed by sulfiredoxin. J Biol Chem. 284, 23364-23374.

Mieyal JJ, Gallogly MM, Qanungo S, Sabens EA, and Shelton MD, Molecular Mechanisms and Clinical Implications of Reversible Protein S-Glutathionylation, invited comprehensive review in Antioxidants & Redox Signaling 10, 1941-1988 (2008).

Shelton, M.D., T.S. Kern & J.J Mieyal, Glutaredoxin Regulates Nuclear Factor kappa-B and Intercellular Adhesion Molecule in Muller Cells: Model of Diabetic Retinopathy, J. Biol. Chem., 282, 12467-12474 (2007).

Gallogly, M.M., D.W. Starke, A.K. Leonberg, S.M. Ospina, and J.J. Mieyal, Kinetic and mechanistic characterization and versatile catalytic properties of mammalian glutaredoxin 2: implications for intracellular roles, Biochemistry 47, 11144 -11157 (2008).

Pai, H.V., D.W. Starke, E.J. Lesnefsky, C.L. Hoppel, and J.J. Mieyal, What is the Functional Significance of the Unique Localization of Glutaredoxin 1 (GRx1) in the Intermembrane Space of Mitochondria? Antioxidants & Redox Signaling, 9, 1-7 (2007).

Gallogly MM, Mieyal JJ., Mechanisms of reversible protein glutathionylation in redox signaling and oxidative stress, Curr Opin Pharmacol. 7, 381-391 (2007).

Qanungo S., D.W. Starke, H.V. Pai, J.J. Mieyal, Nieminen AL., Glutathione Supplementation Potentiates Hypoxic Apoptosis by S-Glutathionylation of p65-NF{kappa}B. J Biol Chem. 282, 18427-18436 (2007).

Jao, S.-C., S. M. English Ospina, C.B. Post, A.J. Berdis, D.W. Starke and J.J. Mieyal (2006), Computational and Mutational Analysis of Human Glutaredoxin (Thioltransferase) - Modeling the Molecular Basis of Catalysis, Biochemistry, 45, 4785-4796.

Asmis R, Wang Y, Xu L, Kisgati M, Begley JG, Mieyal JJ. (2005), A novel thiol oxidation-based mechanism for adriamycin-induced cell injury in human macrophages, FASEB J. 19, 1866-1868.

M.D. Shelton, P.B. Chock & J.J Mieyal , "Glutaredoxin: Role in Reversible Protein S-Glutathionylation and Regulation of Redox Signal Transduction and Protein Translocation," invited Forum Review in Antioxidants & Redox Signaling , K. Nose, Ed., Mary Ann Liebert, Inc., Larchmont, NY, Vol. 7, ps. 346-364 (2005).

Starke, D.W., P.B. Chock, and J.J. Mieyal (2003), Glutathione-Thiyl Radical Scavenging and Transferase Properties of Human Glutaredoxin (Thioltransferase) – Potential Role in Redox Signal Transduction, J.Biol. Chem ., 278, 14607-14613.

Jun Wang, Ephrem Tekle, Hammou Oubrahim, John J. Mieyal , Earl R. Stadtman, and P. Boon Chock (2003), Stable and controllable RNA interference: Investigating the physiological function of glutathionylated actin, Proc. Nat'l. Acad. Sci. U.S.A . 100 , 5103-5106.

C.A. Chrestensen, D.W. Starke, and J.J. Mieyal , Acute cadmium exposure inactivates thioltransferase (glutaredoxin), inhibits intracellular reduction of protein-glutathione mixed disulfides and initiates apoptosis , J. Biol. Chem. 275 , 26556-26565 (2000).

Y. Yang ƒ , S.-C. Jao ƒ , S. Nanduri, D. W. Starke, J J. Mieyal *, and J. Qin*, "Reactivity of the Human Thioltransferase (C7S, C25S, C78S, C82S) Mutant and NMR Solution Structure of its Glutathionyl Mixed Disulfide Intermediate Reflect Catalytic Specificity, Biochemistry 37 , 17145-17156 (1998).

U. Srinivasan, P.A. Mieyal, and J.J. Mieyal , pH Profiles indicative of Rate Limiting Nucleophilic Displacement in Thioltransferase (Glutaredoxin) Catalysis, Biochemistry 36 , 3199-3206 (1997).

D.A. Davis, F.M. Newcomb, D.W. Starke, D. Ott, L. Arthur, J.J. Mieyal , and R.Yarchoan, Glutathionylated Forms of the HIV-1 Protease are Substrates for Human Thioltransferase (Glutaredoxin), an Enzyme also Detected in HIV-1 Virions, J. Biol. Chem. 272 , 25935-25940 (1997).

S.A. Gravina and J.J. Mieyal , "Thioltransferase is a Specific Glutathionyl Mixed Disulfide Oxidoreductase," Biochemistry 32 , 3368-3376 (1993).