| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Protein Science, Vol 7, Issue 5 1233-1244, Copyright © 1998 by Cold Spring Harbor Laboratory Press
ARTICLE |
E. MOSSNER, M. HUBER-WUNDERLICH and R. GLOCKSHUBER
Institut fur Molekularbiologie und Biophysik, Eidgenossische Technische Hochschule Honggerberg, CH-8093 Zurich, Switzerland
Thiol/disulfide oxidoreductases like thioredoxin, glutaredoxin, DsbA, or protein disulfide isomerase (PDI) share the thioredoxin fold and a catalytic disulfide bond with the sequence Cys-Xaa-Xaa-Cys (Xaa corresponds to any amino acid). Despite their structural similarities, the enzymes have very different redox properties, which is reflected by a 100,000-fold difference in the equilibrium constant (K(eq)) with glutathione between the most oxidizing member, DsbA, and the most reducing member, thioredoxin. Here we present a systematic study on a series of variants of thioredoxin from Escherichia coli, in which the Xaa-Xaa dipeptide was exchanged by that of glutaredoxin, PDI, and DsbA. Like the corresponding natural enzymes, all thioredoxin variants proved to be stronger oxidants than the wild-type, with the order wild-type < PDI-type < DsbA-type < glutaredoxin-type. The most oxidizing, glutaredoxin-like variant has a 420-fold decreased value of K(eq), corresponding to an increase in redox potential by 75 mV. While oxidized wild-type thioredoxin is more stable than the reduced form ({Delta}{Delta}G(ox/red) = 16.9 kJ/mol), both redox forms have almost the same stability in the variants. The pH-dependence of the reactivity with the alkylating agent iodoacetamide proved to be the best method to determine the pK(a) value of thioredoxin's nucleophilic active-site thiol (Cys32). A pK(a) of 7.1 was measured for Cys32 in the reduced wild-type. All variants showed a lowered pK(a) of Cys32, with the lowest value of 5.9 for the glutaredoxin-like variant. A correlation of redox potential and the Cys32 pK(a) value could be established on a quantitative level. However, the predicted correlation between the measured {Delta}{Delta}G(ox/red) values and Cys32 pK(a) values was only qualitative.
CiteULike 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Akif, G. Khare, A. K. Tyagi, S. C. Mande, and A. A. Sardesai Functional Studies of Multiple Thioredoxins from Mycobacterium tuberculosis J. Bacteriol., November 1, 2008; 190(21): 7087 - 7095. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Perez-Jimenez, A. P. Wiita, D. Rodriguez-Larrea, P. Kosuri, J. A. Gavira, J. M. Sanchez-Ruiz, and J. M. Fernandez Force-Clamp Spectroscopy Detects Residue Co-evolution in Enzyme Catalysis J. Biol. Chem., October 3, 2008; 283(40): 27121 - 27129. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Roussel, G. Bechade, A. Kriznik, A. Van Dorsselaer, S. Sanglier-Cianferani, G. Branlant, and S. Rahuel-Clermont Evidence for the Formation of a Covalent Thiosulfinate Intermediate with Peroxiredoxin in the Catalytic Mechanism of Sulfiredoxin J. Biol. Chem., August 15, 2008; 283(33): 22371 - 22382. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Masip, D. Klein-Marcuschamer, S. Quan, J. C. A. Bardwell, and G. Georgiou Laboratory Evolution of Escherichia coli Thioredoxin for Enhanced Catalysis of Protein Oxidation in the Periplasm Reveals a Phylogenetically Conserved Substrate Specificity Determinant J. Biol. Chem., January 11, 2008; 283(2): 840 - 848. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Neiers, S. Sonkaria, A. Olry, S. Boschi-Muller, and G. Branlant Characterization of the Amino Acids from Neisseria meningitidis Methionine Sulfoxide Reductase B Involved in the Chemical Catalysis and Substrate Specificity of the Reductase Step J. Biol. Chem., November 2, 2007; 282(44): 32397 - 32405. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Quan, I. Schneider, J. Pan, A. Von Hacht, and J. C. A. Bardwell The CXXC Motif Is More than a Redox Rheostat J. Biol. Chem., September 28, 2007; 282(39): 28823 - 28833. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Gand, M. Antoine, S. Boschi-Muller, and G. Branlant Characterization of the Amino Acids Involved in Substrate Specificity of Methionine Sulfoxide Reductase A J. Biol. Chem., July 13, 2007; 282(28): 20484 - 20491. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Bjur, S. Eriksson-Ygberg, F. Aslund, and M. Rhen Thioredoxin 1 Promotes Intracellular Replication and Virulence of Salmonella enterica Serovar Typhimurium Infect. Immun., September 1, 2006; 74(9): 5140 - 5151. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. D. Ferguson, V. M. Labunskyy, D. E. Fomenko, D. Arac, Y. Chelliah, C. A. Amezcua, J. Rizo, V. N. Gladyshev, and J. Deisenhofer NMR Structures of the Selenoproteins Sep15 and SelM Reveal Redox Activity of a New Thioredoxin-like Family J. Biol. Chem., February 10, 2006; 281(6): 3536 - 3543. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. S. Kulp, E.-M. Frickel, L. Ellgaard, and J. S. Weissman Domain Architecture of Protein-disulfide Isomerase Facilitates Its Dual Role as an Oxidase and an Isomerase in Ero1p-mediated Disulfide Formation J. Biol. Chem., January 13, 2006; 281(2): 876 - 884. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. R. Roberts, Z. A. Wood, T. J. Jonsson, L. B. Poole, and P. A. Karplus Oxidized and synchrotron cleaved structures of the disulfide redox center in the N-terminal domain of Salmonella typhimurium AhpF Protein Sci., September 1, 2005; 14(9): 2414 - 2420. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Huber, D. Boyd, Y. Xia, M. H. Olma, M. Gerstein, and J. Beckwith Use of Thioredoxin as a Reporter To Identify a Subset of Escherichia coli Signal Sequences That Promote Signal Recognition Particle-Dependent Translocation J. Bacteriol., May 1, 2005; 187(9): 2983 - 2991. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Wu, F. Neiers, S. Boschi-Muller, and G. Branlant The N-terminal Domain of PILB from Neisseria meningitidis Is a Disulfide Reductase That Can Recycle Methionine Sulfoxide Reductases J. Biol. Chem., April 1, 2005; 280(13): 12344 - 12350. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Haugstetter, T. Blicher, and L. Ellgaard Identification and Characterization of a Novel Thioredoxin-related Transmembrane Protein of the Endoplasmic Reticulum J. Biol. Chem., March 4, 2005; 280(9): 8371 - 8380. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Moutevelis and J. Warwicker Prediction of pKa and redox properties in the thioredoxin superfamily Protein Sci., October 22, 2004; 13(10): 2744 - 2752. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Neiers, A. Kriznik, S. Boschi-Muller, and G. Branlant Evidence for a New Sub-class of Methionine Sulfoxide Reductases B with an Alternative Thioredoxin Recognition Signature J. Biol. Chem., October 8, 2004; 279(41): 42462 - 42468. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E.-M. Frickel, P. Frei, M. Bouvier, W. F. Stafford, A. Helenius, R. Glockshuber, and L. Ellgaard ERp57 Is a Multifunctional Thiol-Disulfide Oxidoreductase J. Biol. Chem., April 30, 2004; 279(18): 18277 - 18287. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Johansson, C. H. Lillig, and A. Holmgren Human Mitochondrial Glutaredoxin Reduces S-Glutathionylated Proteins with High Affinity Accepting Electrons from Either Glutathione or Thioredoxin Reductase J. Biol. Chem., February 27, 2004; 279(9): 7537 - 7543. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Masip, J. L. Pan, S. Haldar, J. E. Penner-Hahn, M. P. DeLisa, G. Georgiou, J. C. A. Bardwell, and J.-F. Collet An Engineered Pathway for the Formation of Protein Disulfide Bonds Science, February 20, 2004; 303(5661): 1185 - 1189. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. W. Goulding, M. I. Apostol, S. Gleiter, A. Parseghian, J. Bardwell, M. Gennaro, and D. Eisenberg Gram-positive DsbE Proteins Function Differently from Gram-negative DsbE Homologs: A STRUCTURE TO FUNCTION ANALYSIS OF DsbE FROM MYCOBACTERIUM TUBERCULOSIS J. Biol. Chem., January 30, 2004; 279(5): 3516 - 3524. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Schmidt and R. L. Krauth-Siegel Functional and Physicochemical Characterization of the Thioredoxin System in Trypanosoma brucei J. Biol. Chem., November 21, 2003; 278(47): 46329 - 46336. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Antoine, S. Boschi-Muller, and G. Branlant Kinetic Characterization of the Chemical Steps Involved in the Catalytic Mechanism of Methionine Sulfoxide Reductase A from Neisseria meningitidis J. Biol. Chem., November 14, 2003; 278(46): 45352 - 45357. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Zhao, Y. Peng, S.-f. Hao, Z.-h. Zeng, and C.-c. Wang Dimerization by Domain Hybridization Bestows Chaperone and Isomerase Activities J. Biol. Chem., October 31, 2003; 278(44): 43292 - 43298. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. M. Cunnea, A. Miranda-Vizuete, G. Bertoli, T. Simmen, A. E. Damdimopoulos, S. Hermann, S. Leinonen, M. P. Huikko, J.-A. Gustafsson, R. Sitia, et al. ERdj5, an Endoplasmic Reticulum (ER)-resident Protein Containing DnaJ and Thioredoxin Domains, Is Expressed in Secretory Cells or following ER Stress J. Biol. Chem., January 3, 2003; 278(2): 1059 - 1066. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Philipps and R. Glockshuber Randomization of the Entire Active-site Helix alpha 1 of the Thiol-disulfide Oxidoreductase DsbA from Escherichia coli J. Biol. Chem., November 1, 2002; 277(45): 43050 - 43057. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Vinci, J. Couprie, P. Pucci, E. Quemeneur, and M. Moutiez Description of the topographical changes associated to the different stages of the DsbA catalytic cycle Protein Sci., July 1, 2002; 11(7): 1600 - 1612. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Reckenfelderbaumer and R. L. Krauth-Siegel Catalytic Properties, Thiol pK Value, and Redox Potential of Trypanosoma brucei Tryparedoxin J. Biol. Chem., May 10, 2002; 277(20): 17548 - 17555. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Olry, S. Boschi-Muller, M. Marraud, S. Sanglier-Cianferani, A. Van Dorsselear, and G. Branlant Characterization of the Methionine Sulfoxide Reductase Activities of PILB, a Probable Virulence Factor from Neisseria meningitidis J. Biol. Chem., March 29, 2002; 277(14): 12016 - 12022. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. H. Bessette, J. Qiu, J. C. A. Bardwell, J. R. Swartz, and G. Georgiou Effect of Sequences of the Active-Site Dipeptides of DsbA and DsbC on In Vivo Folding of Multidisulfide Proteins in Escherichia coli J. Bacteriol., February 1, 2001; 183(3): 980 - 988. [Abstract] [Full Text]
|
|
|
|
 |
|
 P. H. Bessette, F. Aslund, J. Beckwith, and G. Georgiou Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm PNAS, November 23, 1999; 96(24): 13703 - 13708. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Mossner, M. Huber-Wunderlich, A. Rietsch, J. Beckwith, R. Glockshuber, and F. Aslund Importance of Redox Potential for the in Vivo Function of the Cytoplasmic Disulfide Reductant Thioredoxin from Escherichia coli J. Biol. Chem., September 3, 1999; 274(36): 25254 - 25259. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
