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Molecular and Cellular Biosciences at Wake Forest University

Wake Forest University Graduate School » Molecular and Cellular Biosciences

Leslie B Poole, Ph.D.

Leslie B Poole, Ph.D.

Professor, Biochemistry 

Translational Science Institute
Comprehensive Cancer Center

Organisms living in aerobic environments require mechanisms which prevent or limit damage to cellular components by reactive oxygen species; these species arise from the incomplete reduction of oxygen during respiration or from exposure to external agents such as light, radiation, redox-cycling drugs or stimulated host phagocytes. A variety of enzymatic and nonenzymatic systems have evolved within living organisms to counter such damage. In bacteria such as Salmonella typhimurium and Escherichia coli, expression of a set of antioxidant enzymes is controlled in a coordinate fashion by an oxidation- reduction-sensitive regulatory protein, OxyR. Using a high-expression OxyR mutant of S. typhimurium, a novel enzymatic activity responsible for the NAD(P)H-linked reduction of toxic organic hydroperoxides was discovered. This alkyl hydroperoxide reductase (AhpR), which is the focus of studies in my laboratory, was subsequently shown to be separable into two protein components, designated AhpF and AhpC. AhpF is an FAD-containing protein related to another well- characterized flavoprotein, thioredoxin reductase, and catalyzes the transfer of electrons from NAD(P)H to AhpC. The smaller AhpC protein is without a chromaphoric cofactor and serves directly as the peroxide-reducing component (homologues of AhpC are widespread in biological systems and have been designated "peroxiredoxins"). Our studies of the catalytic mechanism of AhpR indicate that both component proteins operate through cycling of protein- derived cystine disulfides between oxidized (disulfide) and reduced (dithiol) states. Anaerobic titrations of each protein with reductants have confirmed the presence and essentiality of three redox centers in AhpF (one FAD and two disulfide centers) and one redox-active disulfide center per monomer in AhpC (Poole, 1996; Poole, Godzik, et al, 2000). We have also shown that an unusual oxidized cysteine derivative, cysteine-sulfenic acid (Cys-SOH), is generated transiently through direct oxidation of Cys46, one of the two cysteine residues of AhpC, by the hydroperoxide substrate (Ellis & Poole, 1997a & b). Related oxidized cysteine derivatives have been identified in the two other known heme- and metal-independent peroxide reductases (NADH peroxidase and glutathione peroxidases) and may play a role in signal transduction by the OxyR protein itself.

Studies of the structural and chemical bases for the enzymatic functions of AhpF and AhpC involve a wide range of biochemical techniques. Site-directed and PCR-based mutagenesis experiments have been designed to address specific structure-function questions. Characterization of native and mutant proteins involve enzymological techniques, such as spectral titrations and rapid reaction kinetic measurements, protein chemistry methodology to define cysteine content and redox status, and structural work, which includes protein crystallization and analytical ultracentrifugation.


Nelson KJ, Parsonage D, Karplus PA, Poole LB. Evaluating peroxiredoxin sensitivity toward inactivation by peroxide substrates. Methods Enzymol. 2013;527():21-40.


Poole LB, Nelson KJ, Karplus PA. Sulfenic acids and peroxiredoxins in oxidant defense In: Jakob U, Reichmann D, eds. Oxidative stress and redox regulation. Dordrecht: Springer;2013: 85-118.


Keyes JD, Nelson K, Parsonage D, Daniel L, Furdui C, Poole L. Modulation of signaling proteins by reversible cysteine modification [abstract]. FASEB J. 2013;27():993.3.



Qian J, Wani R, Klomsiri C, Poole LB, Tsang AW, Furdui CM. A simple and effective strategy for labeling cysteine sulfenic acid in proteins by utilization of beta-ketoesters as cleavable probes. Chem Commun (Camb). 2012;48(34):4091-4093.


Salsbury FR Jr, Yuan Y, Knaggs MH, Poole LB, Fetrow JS. Structural and electrostatic asymmetry at the active site in typical and atypical peroxiredoxin dimers. J Phys Chem B. 2012;116(23):6832-6843.



Debnath A, Parsonage D, Andrade RM, He C, Cobo ER, Hirata K, Chen S, Garcia-Rivera G, Orozco E, Martinez MB, Gunatilleke SS, Barrios AM, Arkin MR, Poole LB, McKerrow JH, Reed SL. A high-throughput drug screen for Entamoeba histolytica identifies a new lead and target. Nat Med. 2012;18(6):956-960.


Gretes MC, Poole LB, Karplus PA. Peroxiredoxins in parasites. Antioxid Redox Signal. 2012;17(4):608-633.


Salsbury FR Jr, Poole LB, Fetrow JS. Electrostatics of cysteine residues in proteins: parameterization and validation of a simple model. Proteins. 2012;80(11):2583-2591.


Crump KE, Juneau DG, Poole LB, Haas KM, Grayson JM. The reversible formation of cysteine sulfenic acid promotes B-cell activation and proliferation. Eur J Immunol. 2012;42(8):2152-2164.


Perkins A, Gretes MC, Nelson KJ, Poole LB, Karplus PA. Mapping the active site helix-to-strand conversion of CxxxxC peroxiredoxin Q enzymes. Biochemistry. 2012;51(38):7638-7650.


Lang BS, Gorren ACF, Oberdorfer G, Wenzl MV, Furdui CM, Poole LB, Mayer B, Gruber K. Vascular bioactivation of nitroglycerin by aldehyde dehydrogenase-2: reaction intermediates revealed by crystallography and mass spectrometry. J Biol Chem. 2012;287(45):38124-34.


Daniel LW, Klomsiri C, Rogers LC, Nelson KJ, Soito L, King SB, Poole LB.Localized hydrogen peroxide-dependent cysteine oxidation is required for lysophosphatidic acid signaling in ovarian and prostate cancer cells [abstract]. Free Radic Biol Med. 2012;53(Suppl 2):S33.


Michalek RD, Crump KE, Weant AE, Hiltbold EM, Juneau DG, Moon EY, Yu DY, Poole LB, Grayson JM. Peroxiredoxin II regulates effector and secondary memory CD8+ T cell responses. J Virol. 2012;86(24):13629-41.


Nelson KJ, Parsonage D, Van Swearingen AED, Yuan Y, Salsbury FR, Hall A, Karplus PA, Poole LB. Specific residues in peroxiredoxins promote peroxide reactivity through effects on cysteine pKa, transition state stabilization and oligomerization [abstract]. Free Radic Biol Med. 2012;53(Suppl 2):S151.


Poole L, Karplus PA, Nelson K, Parsonage D. Active site and interface communication regulating peroxiredoxin functions [abstract]. Free Radic Biol Med. 2012;53(Suppl 2):S8.


Soito L, Williamson C, Knutson ST, Fetrow JS, Poole LB, Nelson KJ. PREX: PeroxiRedoxin classification indEX, a database of subfamily assignments across the diverse peroxiredoxin family. Nucleic Acids Res. 2011;39(Database issue):D332-7.


Nelson KJ, Knutson ST, Soito L, Klomsiri C, Poole LB, Fetrow JS. Analysis of the peroxiredoxin family: using active-site structure and sequence information for global classification and residue analysis. Proteins. 2011;79(3):947-964.


Klomsiri C, Karplus PA, Poole LB. Cysteine-based redox switches in enzymes. Antioxid Redox Signal. 2011;14(6):1065-1077.


Wani R, Qian J, Yin L, Bechtold E, King SB, Poole LB, Paek E, Tsang AW, Furdui CM. Isoform-specific regulation of Akt by PDGF-induced reactive oxygen species. Proc Natl Acad Sci U S A. 2011;108(26):10550-5.



Hall A, Nelson K, Poole LB, Karplus PA. Structure-based insights into the catalytic power and conformational dexterity of peroxiredoxins. Antioxid Redox Signal. 2011;15(3):795-815.


Qian J, Klomsiri C, Wright MW, King SB, Tsang AW, Poole LB, Furdui CM. Simple synthesis of 1,3-cyclopentanedione derived probes for labeling sulfenic acid proteins. Chem Commun. 2011;47(32):9203-9205.



Kaplan N, Urao N, Furuta E, Kim SJ, Razvi M, Nakamura Y, Mckinney RD, Poole LB, Fukai T, Ushio-Fukai M. Localized cysteine sulfenic acid formation by vascular endothelial growth factor: role in endothelial cell migration and angiogenesis. Free Radic Res. 2011;45(10):1124-1135.


Nelson KJ, Rogers LC, Klomsiri C, Soito L, Poole LB, Daniel LW. Localized hydrogen peroxide-dependent cysteine oxidation is required for lysophosphatidic acid signaling in ovarian and prostate cancer cells [abstract]. Free Radic Biol Med. 2011;51(Suppl 1):S137.


Qian J, Klomsiri C, King SB, Poole LB, Tsang AW, Furdui CM. Simple synthesis of chemical probes for labeling sulfenic acid proteins [abstract]. Free Radic Biol Med. 2011;51(Suppl 1):S20.


Clarke TE, Romanov V, Chirgadze YN, Klomsiri C, Kisselman G, Wu-Brown J, Poole LB, Pai EF, Chirgadze NY. Crystal structure of alkyl hydroperoxidase D like protein PA0269 from Pseudomonas aeruginosa: homology of the AhpD-like structural family. BMC Struct Biol. 2011;11():27.


Reeves SA, Parsonage D, Nelson KJ, Poole LB. Kinetic and thermodynamic features reveal that Escherichia coli BCP is an unusually versatile peroxiredoxin. Biochemistry. 2011;50(41):8970-8981.


Bechtold E, Reisz JA, Klomsiri C, Tsang AW, Wright MW, Poole LB, Furdui CM, King SB. Water-soluble triarylphosphines as biomarkers for protein S-nitrosation. ACS Chem Biol. 2010;5(4):405-414.


Parsonage D, Desrosiers DC, Hazlett KRO, Sun Y, Nelson KJ, Cox DL, Radolf JD, Poole LB. Broad specificity AhpC-like peroxiredoxin and its thioredoxin reductant in the sparse antioxidant defense system of Treponema pallidum. Proc Natl Acad Sci U S A. 2010;107(14):6240-6245.


Oshikawa J, Urao N, Kim HW, Kaplan N, Razvi M, McKinney R, Poole LB, Fukai T, Ushio-Fukai M. Extracellular SOD-derived H2O2 promotes VEGF signaling in caveolae/lipid rafts and post-ischemic angiogenesis in mice. PLoS ONE. 2010;5(4):e10189.


Nelson KJ, Klomsiri C, Codreanu SG, Soito L, Liebler DC, Rogers LC, Daniel LW, Poole LB. Use of dimedone-based chemical probes for sulfenic acid detection methods to visualize and identify labeled proteins. Methods Enzymol. 2010;473():95-115.


Klomsiri C, Nelson KJ, Bechtold E, Soito L, Johnson LC, Lowther WT, Ryu S-E, King SB, Furdui CM, Poole LB. Use of dimedone-based chemical probes for sulfenic acid detection evaluation of conditions affecting probe incorporation into redox-sensitive proteins. Methods Enzymol. 2010;473():77-94.


Parsonage D, Reeves SA, Karplus PA, Poole LB. Engineering of fluorescent reporters into redox domains to monitor electron transfers. Methods Enzymol. 2010;474():1-21.


Yuan Y, Knaggs MH, Poole LB, Fetrow JS, Salsbury FR Jr. Conformational and oligomeric effects on the cysteine pKa of tryparedoxin peroxidase. J Biomol Struct Dyn. 2010;28(1):51-70.


Hall A, Parsonage D, Poole LB, Karplus PA. Structural evidence that peroxiredoxin catalytic power is based on transition-state stabilization. J Mol Biol. 2010;402(1):194-209.


Flaherty NL, Mellilo AA, Rashel M, Poole LB, Melendez JA. Defining the antioxidant determinants of Francisella tularensis virulence [abstract]. Free Radic Biol Med. 2010;49(Suppl 1):S142.


Saunders JA, Rogers LC, Klomsiri C, Poole LB, Daniel LW. Reactive oxygen species mediate lysophosphatidic acid induced signaling in ovarian cancer cells. Free Radic Biol Med. 2010;49(12):2058-2067.


King SB, Poole L, inventors; Wake Forest University Health Sciences, assignee.Sulfenic acid-reactive compounds. United States patent US 7,803,630. 2010 Sept 28. 2010;():.


Hall A, Karplus PA, Poole LB. Typical 2-Cys peroxiredoxins--structures, mechanisms and functions. FEBS J. 2009;276(9):2469-2477.


Hall A, Parsonage D, Horita D, Karplus PA, Poole LB, Barbar E. Redox-dependent dynamics of a dual thioredoxin fold protein: evolution of specialized folds. Biochemistry. 2009;48(25):5984-5993.


Hall A, Sankaran B, Poole LB, Karplus PA. Structural changes common to catalysis in the Tpx peroxiredoxin subfamily. J Mol Biol. 2009;393(4):867-881.


Klomsiri C, Poole LB, Nelson KJ, Rogers L, Klorig EB, Daniel LW, King SB, Furdui CM. Detection and identification of oxidative cysteine modifications in proteins involved in signal transduction pathways [abstract]. Free Radic Biol Med. 2009;47(Suppl 1):S17-S18.


Klorig EB, Reisz JA, Klomsiri C, Wright MW, Day CS, Poole LB, Furdui CM, King B.The reaction of S-nitrosothiols with a water-soluble phosphine gives S-alkyl phosphonium products as potential bio-markers [abstract]. Free Radic Biol Med. 2009;47(Suppl 1):S32.


Kaplan N, Urao N, Razvi M, Nakamura Y, Poole LB, Fukai T, Ushio-Fukai M. ROS-dependent cysteine sulfenic acid formation of IQGAP1 and ERK1/2 in VEGF-induced endothelial cell proliferation and migration [abstract]. Circulation. 2009;120(18 Suppl 2):S1085.


Yamamoto Y, Ritz D, Planson A-G, Jonsson TJ, Faulkner MJ, Boyd D, Beckwith J, Poole LB. Mutant AhpC peroxiredoxins suppress thiol-disulfide redox deficiencies and acquire deglutathionylating activity. Mol Cell. 2008;29(1):36-45.


Poole LB, Nelson KJ. Discovering mechanisms of signaling-mediated cysteine oxidation. Curr Opin Chem Biol. 2008;12(1):18-24.


Salsbury FR Jr, Knutson ST, Poole LB, Fetrow JS. Functional site profiling and electrostatic analysis of cysteines modifiable to cysteine sulfenic acid. Protein Sci. 2008;17(2):299-312.



Parsonage D, Karplus PA, Poole LB. Substrate specificity and redox potential of AhpC, a bacterial peroxiredoxin. Proc Natl Acad Sci U S A. 2008;105(24):8209-8214.


Smith-Pearson PS, Kooshki M, Spitz DR, Poole LB, Zhao W, Robbins ME.Decreasing peroxiredoxin II expression decreases glutathione, alters cell cycle distribution, and sensitizes glioma cells to ionizing radiation and H2O2. Free Radic Biol Med. 2008;45(8):1178-1189.


Nelson KJ, Parsonage D, Hall A, Karplus A, Poole LB. Cysteine pKa values for the bacterial peroxiredoxin AhpC. Biochemistry. 2008;47(48):12860-68.


Hutson SM, Poole LB, Coles S, Conway ME. Redox regulation and trapping sulfenic acid in the peroxide-sensitive human mitochondrial branched chain aminotransferase. Methods Mol Biol. 2008;476():139-152.


Nakamura Y, Kaplan N, Poole LB, Fukai T, Ushio-Fukai M. Detection of cysteine sulfenic acid formation: role in reactive oxygen species-dependent VEGF signaling linked to endothelial migration [abstract]. Circulation. 2008;118(18 Suppl 2):S273.



Allen EE, Diao L, Fetrow JS, John DJ, Loeser RF, Poole LB.The shuffle index and evaluation of models of signal transduction pathways



Poole LB, Klomsiri C, Knaggs SA, Furdui CM, Nelson KJ, Thomas MJ, Fetrow JS, Daniel LW, King SB. Fluorescent and affinity-based tools to detect cysteine sulfenic acid formation in proteins. Bioconjug Chem. 2007;18(6):2004-2017.


Michalek RD, Nelson KJ, Holbrook BC, Yi JS, Stridiron D, Daniel LW, Fetrow JS, King SB, Poole LB, Grayson JM. The requirement of reversible cysteine sulfenic acid formation for T cell activation and function. J Immunol. 2007;179(10):6456-6467.


Poole LB. The catalytic mechanism of peroxiredoxins. Subcell Biochem. 2007;44():61-81.


Klomsiri C, Nelson KJ, Naggs SA, Fetrow JS, King SB, Furdui CM, Poole LB.Cysteine sulfenic acid detection in signal transduction pathways [abstract]. Free Radic Biol Med. 2007;43(Suppl 1):S157.


Nelson KJ, Klomsiri C, Rogers RL, Daniel LW, Poole LB. The role of cysteine sulfenic acid formation during NF-kappa B-mediated signaling [abstract]. Free Radic Biol Med. 2007;43(Suppl 1):S156.


Parsonage D, Nelson KJ, Day AE, Poole LB. Substrate specificity and chemical characterization of a bacterial peroxiredoxin, AhpC [abstract]. Free Radic Biol Med. 2007;43(Suppl 1):S113.


King SB, Poole L, Fetrow J, inventors; Wake Forest University Health Sciences, assignee. Sulfenic acid-reactive compounds and their methods of synthesis. United States patent US 7,294,748. 2007 Nov 13. 2007;():.


Atherosclerosis/thrombosis, biotechnology, cancer/oncogenesis, cell growth, differentiation,d, immunology/allergy/inflammatio, molecular biology/molecular me, structural biology.