HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gromer, S. Articles by Becker, K. Search for Related Content PubMed PubMed Citation Articles by Gromer, S. Articles by Becker, K. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 32, 20096-20101, August 7, 1998

Human Placenta Thioredoxin Reductase
ISOLATION OF THE SELENOENZYME, STEADY STATE KINETICS, AND INHIBITION BY THERAPEUTIC GOLD COMPOUNDS

Stephan Gromer, L. David Arscott^¶, Charles H. Williams Jr.^¶, R. Heiner Schirmer, and Katja Becker

From the  Center of Biochemistry, Heidelberg University, 69120 Heidelberg, Germany, the ^¶ Department of Veterans Affairs Medical Center, University of Michigan, Ann Arbor, Michigan 48105, and the  Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48105

Human thioredoxin reductase is a pyridine nucleotide-disulfide oxidoreductase closely related to glutathione reductase but differing from the latter in having a Cys-SeCys (selenocysteine) sequence as an additional redox center. Because selenoproteins cannot be expressed yet in heterologous systems, we optimized the purification of the protein from placenta with respect to final yield (1-2 mg from one placenta), specific activity (42 units/mg), and selenium content (0.94 ± 0.03 mol/mol subunit). The steady state kinetics showed that the enzyme operates by a ping-pong mechanism; the value of k_cat was 3330 ± 882 min¹, and the K_m values were 18 µM for NADPH and 25 µM for Escherichia coli thioredoxin. The activation energy of the reaction was found to be 53.2 kJ/mol, which allows comparisons of the steady state data with previous pre-steady state measurements. In its physiological, NADPH-reduced form, the enzyme is strongly inhibited by organic gold compounds that are widely used in the treatment of rheumatoid arthritis; for auranofin, the K_i was 4 nM when measured in the presence of 50 µM thioredoxin. At 1000-fold higher concentrations, that is at micromolar levels, the drugs also inhibited human glutathione reductase and the selenoenzyme glutathione peroxidase.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				C. Yan, B. Shieh, P. Reigan, Z. Zhang, M. A. Colucci, A. Chilloux, J. J. Newsome, D. Siegel, D. Chan, C. J. Moody, et al. Potent Activity of Indolequinones against Human Pancreatic Cancer: Identification of Thioredoxin Reductase as a Potential Target Mol. Pharmacol., July 1, 2009; 76(1): 163 - 172. [Abstract] [Full Text] [PDF]

				Q. Cheng, T. Sandalova, Y. Lindqvist, and E. S. J. Arner Crystal Structure and Catalysis of the Selenoprotein Thioredoxin Reductase 1 J. Biol. Chem., February 6, 2009; 284(6): 3998 - 4008. [Abstract] [Full Text] [PDF]

				N. Sturm, Y. Hu, H. Zimmermann, K. Fritz-Wolf, S. Wittlin, S. Rahlfs, and K. Becker Compounds Structurally Related to Ellagic Acid Show Improved Antiplasmodial Activity Antimicrob. Agents Chemother., February 1, 2009; 53(2): 622 - 630. [Abstract] [Full Text] [PDF]

				U. Gundimeda, J. E. Schiffman, D. Chhabra, J. Wong, A. Wu, and R. Gopalakrishna Locally Generated Methylseleninic Acid Induces Specific Inactivation of Protein Kinase C Isoenzymes: RELEVANCE TO SELENIUM-INDUCED APOPTOSIS IN PROSTATE CANCER CELLS J. Biol. Chem., December 12, 2008; 283(50): 34519 - 34531. [Abstract] [Full Text] [PDF]

				A Mukherjee and S G Martin The thioredoxin system: a key target in tumour and endothelial cells Br. J. Radiol., October 1, 2008; 81(Special_Issue_1): S57 - S68. [Abstract] [Full Text] [PDF]

				C. Norez, M. Pasetto, M. C. Dechecchi, E. Barison, C. Anselmi, A. Tamanini, F. Quiri, L. Cattel, P. Rizzotti, F. Dosio, et al. Chemical conjugation of {Delta}F508-CFTR corrector deoxyspergualin to transporter human serum albumin enhances its ability to rescue Cl- channel functions Am J Physiol Lung Cell Mol Physiol, August 1, 2008; 295(2): L336 - L347. [Abstract] [Full Text] [PDF]

				M. Bonilla, A. Denicola, S. V. Novoselov, A. A. Turanov, A. Protasio, D. Izmendi, V. N. Gladyshev, and G. Salinas Platyhelminth Mitochondrial and Cytosolic Redox Homeostasis Is Controlled by a Single Thioredoxin Glutathione Reductase and Dependent on Selenium and Glutathione J. Biol. Chem., June 27, 2008; 283(26): 17898 - 17907. [Abstract] [Full Text] [PDF]

				E.-H. Chew, J. Lu, T. D. Bradshaw, and A. Holmgren Thioredoxin reductase inhibition by antitumor quinols: a quinol pharmacophore effect correlating to antiproliferative activity FASEB J, June 1, 2008; 22(6): 2072 - 2083. [Abstract] [Full Text] [PDF]

				M. Benhar, M. T. Forrester, D. T. Hess, and J. S. Stamler Regulated Protein Denitrosylation by Cytosolic and Mitochondrial Thioredoxins Science, May 23, 2008; 320(5879): 1050 - 1054. [Abstract] [Full Text] [PDF]

				K. Buchholz, R. H. Schirmer, J. K. Eubel, M. B. Akoachere, T. Dandekar, K. Becker, and S. Gromer Interactions of Methylene Blue with Human Disulfide Reductases and Their Orthologues from Plasmodium falciparum Antimicrob. Agents Chemother., January 1, 2008; 52(1): 183 - 191. [Abstract] [Full Text] [PDF]

				T. E. Tipple, S. E. Welty, L. K. Rogers, T. N. Hansen, Y.-E. Choi, J. P. Kehrer, and C. V. Smith Thioredoxin-Related Mechanisms in Hyperoxic Lung Injury in Mice Am. J. Respir. Cell Mol. Biol., October 1, 2007; 37(4): 405 - 413. [Abstract] [Full Text] [PDF]

				J. P. Gray, D. E. Heck, V. Mishin, P. J. S. Smith, J.-Y. Hong, M. Thiruchelvam, D. A. Cory-Slechta, D. L. Laskin, and J. D. Laskin Paraquat Increases Cyanide-insensitive Respiration in Murine Lung Epithelial Cells by Activating an NAD(P)H:Paraquat Oxidoreductase: IDENTIFICATION OF THE ENZYME AS THIOREDOXIN REDUCTASE J. Biol. Chem., March 16, 2007; 282(11): 7939 - 7949. [Abstract] [Full Text] [PDF]

				N. Nagahara, T. Yoshii, Y. Abe, and T. Matsumura Thioredoxin-dependent Enzymatic Activation of Mercaptopyruvate Sulfurtransferase: AN INTERSUBUNIT DISULFIDE BOND SERVES AS A REDOX SWITCH FOR ACTIVATION J. Biol. Chem., January 19, 2007; 282(3): 1561 - 1569. [Abstract] [Full Text] [PDF]

				P. B. Cassidy, K. Edes, C. C. Nelson, K. Parsawar, F.A. Fitzpatrick, and P. J. Moos Thioredoxin reductase is required for the inactivation of tumor suppressor p53 and for apoptosis induced by endogenous electrophiles Carcinogenesis, December 1, 2006; 27(12): 2538 - 2549. [Abstract] [Full Text] [PDF]

				K. M. Massimine, M. T. McIntosh, L. T. Doan, C. E. Atreya, S. Gromer, W. Sirawaraporn, D. A. Elliott, K. A. Joiner, R. H. Schirmer, and K. S. Anderson Eosin B as a Novel Antimalarial Agent for Drug-Resistant Plasmodium falciparum. Antimicrob. Agents Chemother., September 1, 2006; 50(9): 3132 - 3141. [Abstract] [Full Text] [PDF]

				A. V. Lobanov, S. Gromer, G. Salinas, and V. N. Gladyshev Selenium metabolism in Trypanosoma: characterization of selenoproteomes and identification of a Kinetoplastida-specific selenoprotein Nucleic Acids Res., September 1, 2006; 34(14): 4012 - 4024. [Abstract] [Full Text] [PDF]

				N. Cenas, S. Prast, H. Nivinskas, J. Sarlauskas, and E. S. J. Arner Interactions of Nitroaromatic Compounds with the Mammalian Selenoprotein Thioredoxin Reductase and the Relation to Induction of Apoptosis in Human Cancer Cells J. Biol. Chem., March 3, 2006; 281(9): 5593 - 5603. [Abstract] [Full Text] [PDF]

				K.-R. Erlemann, J. Rokach, and W. S. Powell Oxidative Stress Stimulates the Synthesis of the Eosinophil Chemoattractant 5-Oxo-6,8,11,14-eicosatetraenoic Acid by Inflammatory Cells J. Biol. Chem., September 24, 2004; 279(39): 40376 - 40384. [Abstract] [Full Text] [PDF]

				M. K. Yu, P. J. Moos, P. Cassidy, M. Wade, and F. A. Fitzpatrick Conditional Expression of 15-Lipoxygenase-1 Inhibits the Selenoenzyme Thioredoxin Reductase: MODULATION OF SELENOPROTEINS BY LIPOXYGENASE ENZYMES J. Biol. Chem., July 2, 2004; 279(27): 28028 - 28035. [Abstract] [Full Text] [PDF]

				N. Cenas, H. Nivinskas, Z. Anusevicius, J. Sarlauskas, F. Lederer, and E. S. J. Arner Interactions of Quinones with Thioredoxin Reductase: A CHALLENGE TO THE ANTIOXIDANT ROLE OF THE MAMMALIAN SELENOPROTEIN J. Biol. Chem., January 23, 2004; 279(4): 2583 - 2592. [Abstract] [Full Text] [PDF]

				S. Gromer, L. Johansson, H. Bauer, L. D. Arscott, S. Rauch, D. P. Ballou, C. H. Williams Jr., R. H. Schirmer, and E. S. J. Arner Active sites of thioredoxin reductases: Why selenoproteins? PNAS, October 28, 2003; 100(22): 12618 - 12623. [Abstract] [Full Text] [PDF]

				H. Bauer, V. Massey, L. D. Arscott, R. H. Schirmer, D. P. Ballou, and C. H. Williams Jr. The Mechanism of High Mr Thioredoxin Reductase from Drosophila melanogaster J. Biol. Chem., August 29, 2003; 278(35): 33020 - 33028. [Abstract] [Full Text] [PDF]

				G. G. J. M. Kuiper, W. Klootwijk, and T. J. Visser Substitution of Cysteine for Selenocysteine in the Catalytic Center of Type III Iodothyronine Deiodinase Reduces Catalytic Efficiency and Alters Substrate Preference Endocrinology, June 1, 2003; 144(6): 2505 - 2513. [Abstract] [Full Text] [PDF]

				K. Anestal and E. S. J. Arner Rapid Induction of Cell Death by Selenium-compromised Thioredoxin Reductase 1 but Not by the Fully Active Enzyme Containing Selenocysteine J. Biol. Chem., April 25, 2003; 278(18): 15966 - 15972. [Abstract] [Full Text] [PDF]

				A. Agorio, C. Chalar, S. Cardozo, and G. Salinas Alternative mRNAs Arising from Trans-splicing Code for Mitochondrial and Cytosolic Variants of Echinococcus granulosus Thioredoxin Glutathione Reductase J. Biol. Chem., April 4, 2003; 278(15): 12920 - 12928. [Abstract] [Full Text] [PDF]

				P. J. Moos, K. Edes, P. Cassidy, E. Massuda, and F. A. Fitzpatrick Electrophilic Prostaglandins and Lipid Aldehydes Repress Redox-sensitive Transcription Factors p53 and Hypoxia-inducible Factor by Impairing the Selenoprotein Thioredoxin Reductase J. Biol. Chem., January 3, 2003; 278(2): 745 - 750. [Abstract] [Full Text] [PDF]

				F. Missirlis, J. K. Ulschmid, M. Hirosawa-Takamori, S. Gronke, U. Schafer, K. Becker, J. P. Phillips, and H. Jackle Mitochondrial and Cytoplasmic Thioredoxin Reductase Variants Encoded by a Single Drosophila Gene Are Both Essential for Viability J. Biol. Chem., March 22, 2002; 277(13): 11521 - 11526. [Abstract] [Full Text] [PDF]

				S. Gromer and J. H. Gross Methylseleninate Is a Substrate Rather Than an Inhibitor of Mammalian Thioredoxin Reductase. IMPLICATIONS FOR THE ANTITUMOR EFFECTS OF SELENIUM J. Biol. Chem., March 15, 2002; 277(12): 9701 - 9706. [Abstract] [Full Text] [PDF]

				Z. Gregus, A. Gyurasics, and I. Csanaky Effects of Arsenic-, Platinum-, and Gold-Containing Drugs on the Disposition of Exogenous Selenium in Rats Toxicol. Sci., September 1, 2000; 57(1): 22 - 31. [Abstract] [Full Text] [PDF]

				P. C. Billings, D. J. Herrick, P. S. Howard, U. Kucich, B. N. Engelsberg, and J. Rosenbloom Expression of beta ig-h3 by Human Bronchial Smooth Muscle Cells . Localization to the Extracellular Matrix and Nucleus Am. J. Respir. Cell Mol. Biol., March 1, 2000; 22(3): 352 - 359. [Abstract] [Full Text]

				H. E. Ganther Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase Carcinogenesis, September 1, 1999; 20(9): 1657 - 1666. [Abstract] [Full Text] [PDF]

				Q.-A. Sun, Y. Wu, F. Zappacosta, K.-T. Jeang, B. J. Lee, D. L. Hatfield, and V. N. Gladyshev Redox Regulation of Cell Signaling by Selenocysteine in Mammalian Thioredoxin Reductases J. Biol. Chem., August 27, 1999; 274(35): 24522 - 24530. [Abstract] [Full Text] [PDF]

				L. Zhong and A. Holmgren Essential Role of Selenium in the Catalytic Activities of Mammalian Thioredoxin Reductase Revealed by Characterization of Recombinant Enzymes with Selenocysteine Mutations J. Biol. Chem., June 9, 2000; 275(24): 18121 - 18128. [Abstract] [Full Text] [PDF]

				S. M. Kanzok, R. H. Schirmer, I. Turbachova, R. Iozef, and K. Becker The Thioredoxin System of the Malaria Parasite Plasmodium falciparum. GLUTATHIONE REDUCTION REVISITED J. Biol. Chem., December 15, 2000; 275(51): 40180 - 40186. [Abstract] [Full Text] [PDF]

				L. Zhong, E. S. J. Arner, and A. Holmgren Structure and mechanism of mammalian thioredoxin reductase: The active site is a redox-active selenolthiol/selenenylsulfide formed from the conserved cysteine-selenocysteine sequence PNAS, May 23, 2000; 97(11): 5854 - 5859. [Abstract] [Full Text] [PDF]