Comparative modeling of thioredoxin glutathione reductase from Schistosoma mansoni: a multifunctional target for antischistosomal therapy.

Schistosoma mansoni, a trematode parasite, which causes schistosomiasis and affects more than 200 million people worldwide, lives in an aerobic environment and therefore needs an effective redox mechanism for surviving reactive oxygen species from its host. Although, the host has two different redox systems: glutaredoxin and thioredoxin, the parasite has only one unique multifunctional enzyme, thioredoxin glutathione reductase (TGR) involving a fusion of two proteins, glutaredoxin (Grx) and thioredoxin reductase (TR), for performing all the redox activities. This dependence of S. mansoni on a single protein, TGR, for its protection from oxidative stress, makes it a promising drug target. Here, we describe a suitably validated, homology model for S. mansoni TGR (SmTGR), developed using both TR and Grx templates, functionally complete in the dimeric form with cofactors NADP(H) and FAD. Comparative analysis of substrate and inhibitor binding pockets of our model with crystal structures of parent TR as well as with that of glutathione reductase (GR), which is an essential component of the Grx system, appears to provide greater insight into the functioning of TGR. This also augments recent observations reported on the basis of X-ray structure data on SmTGR monomer lacking the C-terminal selenocysteine tail.

[1]  J. Daub,et al.  Helminth genome analysis: the current status of the filarial and schistosome genome projects , 1999, Parasitology.

[2]  G. Mendoza-Hernández,et al.  Purification, characterization and kinetic properties of the multifunctional thioredoxin-glutathione reductase from Taenia crassiceps metacestode (cysticerci). , 2004, Molecular and biochemical parasitology.

[3]  V. Gladyshev,et al.  Selenoprotein oxidoreductase with specificity for thioredoxin and glutathione systems , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[4]  L. Chitsulo,et al.  Focus: Schistosomiasis , 2004, Nature Reviews Microbiology.

[5]  C. Czerkinsky,et al.  Current status and future prospects for a vaccine against schistosomiasis , 2004, Expert review of vaccines.

[6]  R. Schirmer,et al.  Substitution of the thioredoxin system for glutathione reductase in Drosophila melanogaster. , 2001, Science.

[7]  R. Judson Genetic Algorithms and Their Use in Chemistry , 2007 .

[8]  J Kuriyan,et al.  Crystal Structure of Escherichia coli Thioredoxin Reductase Refined at 2 Å Resolution , 1994 .

[9]  S. Novoselov,et al.  Mammalian Selenoprotein Thioredoxin-glutathione Reductase , 2005, Journal of Biological Chemistry.

[10]  S. Novoselov,et al.  Reaction mechanism and regulation of mammalian thioredoxin/glutathione reductase. , 2005, Biochemistry.

[11]  K. Soda,et al.  Selenocysteine. , 2020, Methods in enzymology.

[12]  Heather M. Alger,et al.  The disulfide redox system of Schistosoma mansoni and the importance of a multifunctional enzyme, thioredoxin glutathione reductase. , 2002, Molecular and biochemical parasitology.

[13]  C. Jin,et al.  Structural insight into poplar glutaredoxin C1 with a bridging iron-sulfur cluster at the active site. , 2006, Biochemistry.

[14]  A. Holmgren,et al.  Thiol redox control via thioredoxin and glutaredoxin systems. , 2005, Biochemical Society transactions.

[15]  V. Yardley,et al.  A fluoro analogue of the menadione derivative 6-[2'-(3'-methyl)-1',4'-naphthoquinolyl]hexanoic acid is a suicide substrate of glutathione reductase. Crystal structure of the alkylated human enzyme. , 2006, Journal of the American Chemical Society.

[16]  A. Miele,et al.  Glutathione reductase and thioredoxin reductase at the crossroad: The structure of Schistosoma mansoni thioredoxin glutathione reductase , 2008, Proteins.

[17]  V. Gladyshev,et al.  Crystal structures of oxidized and reduced mitochondrial thioredoxin reductase provide molecular details of the reaction mechanism. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[18]  C. Pueyo,et al.  Absolute Gene Expression Patterns of Thioredoxin and Glutaredoxin Redox Systems in Mouse* , 2003, Journal of Biological Chemistry.

[19]  A. Sali,et al.  Modeller: generation and refinement of homology-based protein structure models. , 2003, Methods in enzymology.

[20]  Anastassis Perrakis,et al.  Developments in the CCP4 molecular-graphics project. , 2004, Acta crystallographica. Section D, Biological crystallography.

[21]  G. Schneider,et al.  Three-dimensional structure of a mammalian thioredoxin reductase: Implications for mechanism and evolution of a selenocysteine-dependent enzyme , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[22]  P. Karplus,et al.  Refined structure of glutathione reductase at 1.54 A resolution. , 1987, Journal of molecular biology.

[23]  K. Fritz-Wolf,et al.  The structure of human thioredoxin reductase 1 provides insights into C-terminal rearrangements during catalysis. , 2007, Journal of molecular biology.

[24]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[25]  P Willett,et al.  Use of techniques derived from graph theory to compare secondary structure motifs in proteins. , 1990, Journal of molecular biology.

[26]  C. Fernández,et al.  Linked thioredoxin-glutathione systems in platyhelminths. , 2004, Trends in parasitology.

[27]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[28]  Elias S. J. Arnér,et al.  Structure and mechanism of mammalian thioredoxin reductase: the active site is a redox-active selenolthiol/selenenylsulfide formed from the conserved cysteine-selenocysteine sequence. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  A. W. Schüttelkopf,et al.  PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. , 2004, Acta crystallographica. Section D, Biological crystallography.

[30]  P. Karplus,et al.  Substrate binding and catalysis by glutathione reductase as derived from refined enzyme: substrate crystal structures at 2 A resolution. , 1994, Journal of molecular biology.

[31]  G. Schulz,et al.  Structure of glutathione reductase from escherichia coli at 1.86 Å resolution: Comparison with the enzyme from human erythrocytes , 1994, Protein science : a publication of the Protein Society.

[32]  T C Stadtman,et al.  Selenocysteine, identified as the penultimate C-terminal residue in human T-cell thioredoxin reductase, corresponds to TGA in the human placental gene. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[33]  K. Fritz-Wolf,et al.  Truncated mutants of human thioredoxin reductase 1 do not exhibit glutathione reductase activity , 2006, FEBS letters.

[34]  E. Davioud‐Charvet,et al.  Thioredoxin Glutathione Reductase from Schistosoma mansoni: An Essential Parasite Enzyme and a Key Drug Target , 2007, PLoS medicine.

[35]  P. Karplus,et al.  Kinetics and Crystallographic Analysis of Human Glutathione Reductase in Complex with a Xanthene Inhibitor (*) , 1996, The Journal of Biological Chemistry.

[36]  K Schulten,et al.  VMD: visual molecular dynamics. , 1996, Journal of molecular graphics.

[37]  Laxmikant V. Kalé,et al.  Scalable molecular dynamics with NAMD , 2005, J. Comput. Chem..

[38]  T. Darden,et al.  Particle mesh Ewald: An N⋅log(N) method for Ewald sums in large systems , 1993 .

[39]  V. Gladyshev,et al.  Redox Regulation of Cell Signaling by Selenocysteine in Mammalian Thioredoxin Reductases* , 1999, The Journal of Biological Chemistry.

[40]  S. Nanduri,et al.  Reactivity of the human thioltransferase (glutaredoxin) C7S, C25S, C78S, C82S mutant and NMR solution structure of its glutathionyl mixed disulfide intermediate reflect catalytic specificity. , 1998, Biochemistry.

[41]  Yanfeng Yang,et al.  Crystal structure of thioltransferase at 2.2 Å resolution , 1995, Protein science : a publication of the Protein Society.

[42]  S. Everse,et al.  Structural and biochemical studies reveal differences in the catalytic mechanisms of mammalian and Drosophila melanogaster thioredoxin reductases. , 2007, Biochemistry.

[43]  I. Schlichting,et al.  Ajoene is an inhibitor and subversive substrate of human glutathione reductase and Trypanosoma cruzi trypanothione reductase: crystallographic, kinetic, and spectroscopic studies. , 1999, Journal of medicinal chemistry.

[44]  W. L. Jorgensen,et al.  Comparison of simple potential functions for simulating liquid water , 1983 .