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:
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).