John J. Mieyal, Ph.D.
Professor of Pharmacology and Vice Chair
Phone: (216) 368-3384
Fax: (216) 368-3395
E-mail: john.mieyal@case.edu
Wood RT-300
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.
Selected References:
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.
Nulton-Persson, A.C., D.W. Starke, J.J. Mieyal , and L.I. Szweda (2003), Reversible Inactivation of &alpha-Ketoglutarate Dehydrogenase in Response to Alterations in Mitochondrial Glutathione Status, Biochemistry , 42, 4235-4242.
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.
Biaglow, J.E., I.S. Ayene, C. I. Koch, J. Donahue, T.D. Stomato, J.J. Mieyal, and S.W. Tuttle (2003), Radiation Response of Cells During Altered Protein Thiol Redox, Radiation Research , 159 , 484-494.
J.E. Biaglow, I. Lee, J. Donahue, K. Held, J.J. Mieyal , M. Dewhirst, and S. Tuttle, Glutathione Depletion or Radiation Treatment Alters Respiration and Induces Apoptosis in R3230AC Mammary Carcinoma, invited manuscript for Advances in Experimental Biology and Medicine, Oxygen Transport to Tissue XXIV, J.F. Dunn & H.M. Swartz, Eds., Kluwer Academic /Plenum Publishers, Volume 530, Chapter 15, ps. 153-164 (2003).
J. Ehrhart, M. Gluck, J.J. Mieyal, and G.D. Zeevalk, Functional Glutaredoxin (Thioltransferase) Activity in Rat Brain and Liver Mitochondria, Parkinsonism and Related Disorders . 8 (6), 395-400 (2002)
J. Wang, E. S. Boja, W. Tan, E. Tekel, H. Fales, S.M. English, J.J. Mieyal and P. B. Chock, Reversible Glutathionylation Regulates Actin Polymerization in A431 Cells, J. Biol. Chem. rapid communication 276 , 47763-47766 (2001).
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).
J.E. Biaglow, J. Donahue, S.W. Tuttle, K. Held, C.A. Chrestensen, and J.J. Mieyal , "A Method for Measuring Disulfide Reduction in Cultured Mammalian Cells: The Relative Roles of Glutathione-Dependent and Glutathione-Independent Enzyme Systems," Analytical Biochem . 281 , 77-86 (2000).
Balijepalli, S., Tirumalai, P.S., Swamy, K.V., Boyd, M.R., Mieyal, J.J. , Ravindranath, V. 1999. Rat brain thioltransferase: regional distribution, immunological characterization, and localization by fluorescent in situ hybridization. J. Neurochem. 72:1170-1178.
S.Bandyopadhyay,.W. D Starke, J.J. Mieyal and R.M. Gronostajski, "Thioltransferase (glutaredoxin) reactivates the DNA-binding activity of oxidation-inactivated Nuclear Factor I," J. Biol. Chem. 273 , 392-397 (1998).
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.W. Starke, Y. Chen, C.P. Bapna, E.J. Lesnefsky, and J.J. Mieyal , " Sensitivity of Protein Sulfhydryl Repair Enzymes to Oxidative Stress," Free Radical Biol. Med . 23 , 373-384 (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).
J.J. Mieyal, S.A. Gravina, P.A. Mieyal, U. Srinivasan, and D.W. Starke, Glutathionyl Specificity of Thioltransferases: Mechanistic and Physiological Implications, invited review in Biothiols in Health and Disease, L. Packer & E. Cadenas, Eds., Marcel Dekker, Inc., New York, Chapter 14, ps. 305-372 (1995).
C.A. Chrestensen, C.B. Eckman, D.W. Starke, & J.J. Mieyal , "Cloning, Expression, and Characterization of Human Thioltransferase (Glutaredoxin) in E. coli, " FEBS Lett . 134 , 25-28 (1995).
J.J. Mieyal and D.W. Starke, "Hemoglobin-catalyzed Hydroxylation and Dealkylation Reactions," invited review in Methods of Enzymology , Chapter 38, Vol. 231, ps. 573-598 Academic Press, New York (1994).
J. Talafous, L.M. Sayre, J.J. Mieyal, and G. Klopman, "META: A Dictionary Model of Mammalian Xenobiotic Metabolism," J. Chem. Inform. Computer Sci ., 34 , 1326- 1333 (1994).
S.A. Gravina and J.J. Mieyal , "Thioltransferase is a Specific Glutathionyl Mixed Disulfide Oxidoreductase," Biochemistry 32 , 3368-3376 (1993).