GLUTATHIONE TRANSFERASES-DEFINING STRUCTURE & FUNCTION

Summary

Principal Investigator: R N Armstrong
Affiliation: Vanderbilt University
Country: USA
Abstract: DESCRIPTION (provided by applicant): One of the major challenges in biochemistry is to understand the functional genomics of organisms. It is a staggering problem when one considers the fact that about 40% of the genes in of one of the best-understood organisms, Escherichia coli, are not experimentally defined. The organism colonizes the human intestine shortly after birth and is, along with its close relatives such as Salmonella, Shigella and Yersinia, a pathogen that is a significant source of human disease. Glutathione (GSH) is the predominant redox active thiol in most aerobic organisms where it plays a fundamental role in metabolic, catabolic and redox chemistry. GSH transferases are enzymes that participate in this chemistry by adding GSH to electrophilic acceptors. The E. coli genome harbors genes encoding nine glutathione (GSH) transferase homologues. Amazingly, only one gene has a reasonably well-defined function and it does NOT encode a GSH transferase but rather stringent starvation protein A, SspA, a transcription factor. Under aerobic conditions, GSH is the predominate thiol in E. coli. However it is largely converted to glutathionylspermidine (GspSH) under anaerobic conditions by glutathionylspermidine synthetase/amidase (GSS). The broad, long-term objectives of this project are to understand the GSH/GspSH homeostasis in E. coli and the biological activities and roles of chromosomally encoded GSH transferase homologues in E. coli. Ten interrelated proteins are the subject of this investigation. They include the eight canonical GSH transferase homologues, YliJ, YncG, Gst, YfcF, YfcG, YghU, SspA, and YibF as well as GSS, and the membrane-bound GSH transferase, YecN. The research plan includes three specific aims. The first aim is to understand the mechanism and regulation of GSH/GspSH homeostasis in E. coli. The second aim is to define the biological functions of the eight canonical GSH transferase homologues in E. coli. The third aim is to determine the biological role of the single membrane-bound GSH transferase homologue. These aims will be achieved by a multidisciplinary approach that includes: (i) analysis of genome context;(ii) phenotypic responses to gene knockouts;(iii) response of gene expression to growth conditions and stress;(iv) a search for protein partners;(v) determination of X-ray crystal structures of the proteins;(vi) functional and kinetic characterizations of the proteins. PUBLIC HEALTH RELEVANCE: Escherichia coli is a member of the Enterobacteriaceae family that includes several intestinal pathogens such as Salmonella, Shigella and Yersinia. The bacterium typically colonizes the human intestine shortly after birth where it remains the predominant facultative microorganism. Understanding the biochemistry of E. coli is fundamental to understanding its pathogenesis.
Funding Period: ----------------1982 - ---------------2012-
more information: NIH RePORT

Top Publications

  1. ncbi Double mutation at the subunit interface of glutathione transferase rGSTM1-1 results in a stable, folded monomer
    Lawrence C Thompson
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 0146, USA
    Biochemistry 45:2267-73. 2006
  2. pmc Structural and chemical aspects of resistance to the antibiotic fosfomycin conferred by FosB from Bacillus cereus
    Matthew K Thompson
    Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, United States
    Biochemistry 52:7350-62. 2013
  3. pmc His86 from the N-terminus of frataxin coordinates iron and is required for Fe-S cluster synthesis
    Leslie E Gentry
    Department of Chemistry, The University of Alabama, Tuscaloosa, Alabama 35487 0336, United States
    Biochemistry 52:6085-96. 2013
  4. pmc Synthesis of bacillithiol and the catalytic selectivity of FosB-type fosfomycin resistance proteins
    Alexander P Lamers
    Department of Chemistry, Vanderbilt University, Vanderbilt Institute of Chemical Biology, Nashville, Tennessee 37235, USA
    Org Lett 14:5207-9. 2012
  5. pmc Structure and Function of the Genomically Encoded Fosfomycin Resistance Enzyme, FosB, from Staphylococcus aureus
    Matthew K Thompson
    Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, United States
    Biochemistry 53:755-65. 2014
  6. pmc Characterization of the binding of 8-anilinonaphthalene sulfonate to rat class Mu GST M1-1
    Nichole Kinsley
    Protein Structure Function Research Unit, School of Molecular and Cell Biology, University of the Wiwatersrand, Johannesburg 2050, South Africa
    Biophys Chem 137:100-4. 2008
  7. ncbi Structural elements involved in proton translocation by cytochrome c oxidase as revealed by backbone amide hydrogen-deuterium exchange of the E286H mutant
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University, Nashville, Tennessee 37232 0146, USA
    Biochemistry 47:73-83. 2008
  8. pmc Mapping protein dynamics in catalytic intermediates of the redox-driven proton pump cytochrome c oxidase
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, TN 37232 0146, USA
    Proc Natl Acad Sci U S A 103:15398-403. 2006
  9. pmc Conformational dynamics in the F/G segment of CYP51 from Mycobacterium tuberculosis monitored by FRET
    Galina I Lepesheva
    Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232 0146, USA
    Arch Biochem Biophys 464:221-7. 2007
  10. ncbi Location of substrate binding sites within the integral membrane protein microsomal glutathione transferase-1
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 0146, USA
    Biochemistry 46:2812-22. 2007

Scientific Experts

  • Richard N Armstrong
  • Laura S Busenlehner
  • Matthew K Thompson
  • Edward B Prage
  • Ralf Morgenstern
  • Nina V Stourman
  • Paul D Cook
  • Mary E Keithly
  • Lawrence C Thompson
  • Joel Harp
  • Jane E Ladner
  • Johan Alander
  • Kevin L Jagessar
  • Leslie E Gentry
  • Gary A Sulikowski
  • Anita M Preininger
  • Donald F Stec
  • Alexander P Lamers
  • Kwangho Kim
  • Per Johan Jakobsson
  • Joel M Harp
  • Megan C Wadington
  • Hans Hebert
  • Peter J Holm
  • Nichole Kinsley
  • Galina I Lepesheva
  • Peter Brzezinski
  • Heini W Dirr
  • Eric P Skaar
  • Michael C Goodman
  • Neal D Hammer
  • Russell Timkovich
  • Reece Doughty
  • Matthew A Thacker
  • Heidi E Hamm
  • James A Gilbert
  • Kelly M Hines
  • Ali I Kaya
  • Markus W Voehler
  • Megan C Branch
  • Matthew R Schaab
  • Sven Christian Pawelzik
  • Richard Svensson
  • William J Griffiths
  • Pascal Gerbaux
  • Johan Lengqvist
  • Robert H H van den Heuvel
  • Ida Namslauer
  • Salerwe Mosebi
  • Gisela Brändén
  • Yasien Sayed
  • Damjana Rozman
  • Michael R Waterman
  • Matej Seliskar
  • Charles G Knutson
  • Caroline Jegerscöhld
  • Priyaranjan Bhakat
  • Simona G Codreanu
  • Gary L Gilliland
  • John Walters
  • James F Parsons
  • Lina Salomonsson
  • Jonathan Burke

Detail Information

Publications17

  1. ncbi Double mutation at the subunit interface of glutathione transferase rGSTM1-1 results in a stable, folded monomer
    Lawrence C Thompson
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 0146, USA
    Biochemistry 45:2267-73. 2006
    ..The H/D exchange data reveal a possible molecular basis for the folding cooperativity observed between domains 1 and 2...
  2. pmc Structural and chemical aspects of resistance to the antibiotic fosfomycin conferred by FosB from Bacillus cereus
    Matthew K Thompson
    Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, United States
    Biochemistry 52:7350-62. 2013
    ..The fact that Zn(2+) is an inhibitor of FosB(Bc) was used to obtain a ternary complex structure of the enzyme with both fosfomycin and L-Cys bound...
  3. pmc His86 from the N-terminus of frataxin coordinates iron and is required for Fe-S cluster synthesis
    Leslie E Gentry
    Department of Chemistry, The University of Alabama, Tuscaloosa, Alabama 35487 0336, United States
    Biochemistry 52:6085-96. 2013
    ..Our data suggest that iron coordination to frataxin may be significant to the Fe-S cluster biosynthesis pathway in mitochondria. ..
  4. pmc Synthesis of bacillithiol and the catalytic selectivity of FosB-type fosfomycin resistance proteins
    Alexander P Lamers
    Department of Chemistry, Vanderbilt University, Vanderbilt Institute of Chemical Biology, Nashville, Tennessee 37235, USA
    Org Lett 14:5207-9. 2012
    ..However, the metal-ion selectivity is less distinct with FosB from Bacillus subtilis, Bacillus anthracis, or Bacillus cereus...
  5. pmc Structure and Function of the Genomically Encoded Fosfomycin Resistance Enzyme, FosB, from Staphylococcus aureus
    Matthew K Thompson
    Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, United States
    Biochemistry 53:755-65. 2014
    ..This work identifies FosB as a primary fosfomycin-modifying pathway of S. aureus and establishes the enzyme as a potential therapeutic target for increased efficacy of fosfomycin against the pathogen. ..
  6. pmc Characterization of the binding of 8-anilinonaphthalene sulfonate to rat class Mu GST M1-1
    Nichole Kinsley
    Protein Structure Function Research Unit, School of Molecular and Cell Biology, University of the Wiwatersrand, Johannesburg 2050, South Africa
    Biophys Chem 137:100-4. 2008
    ..The binding properties are compared with those of other GSTs and ANS-binding proteins...
  7. ncbi Structural elements involved in proton translocation by cytochrome c oxidase as revealed by backbone amide hydrogen-deuterium exchange of the E286H mutant
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University, Nashville, Tennessee 37232 0146, USA
    Biochemistry 47:73-83. 2008
    ..In the E286H mutant, the proton gate does not function properly and the exit channel is unresponsive. These results provide new insight into the structure and mechanism of proton translocation by CytcO...
  8. pmc Mapping protein dynamics in catalytic intermediates of the redox-driven proton pump cytochrome c oxidase
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, TN 37232 0146, USA
    Proc Natl Acad Sci U S A 103:15398-403. 2006
    ....
  9. pmc Conformational dynamics in the F/G segment of CYP51 from Mycobacterium tuberculosis monitored by FRET
    Galina I Lepesheva
    Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232 0146, USA
    Arch Biochem Biophys 464:221-7. 2007
    ..The results confirm (1) functional importance of conformational motions in the MTCYP51 F/G segment and (2) applicability of FRET to monitor them in solution...
  10. ncbi Location of substrate binding sites within the integral membrane protein microsomal glutathione transferase-1
    Laura S Busenlehner
    Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 0146, USA
    Biochemistry 46:2812-22. 2007
    ..Such structural changes in MGST1 are essential for activation of the enzyme and are important for its biological function...
  11. ncbi 2-Hydroxychromene-2-carboxylic acid isomerase: a kappa class glutathione transferase from Pseudomonas putida
    Lawrence C Thompson
    Department of Biochemistry, Center in Molecular Toxicology, and Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 0146, USA
    Biochemistry 46:6710-22. 2007
    ..78 A) to C7 of HCCA. Taken together, the results suggest that the isomerization reaction involves a short-lived covalent adduct between the sulfur of GSH and C7 of the substrate...
  12. pmc Observation of two modes of inhibition of human microsomal prostaglandin E synthase 1 by the cyclopentenone 15-deoxy-Δ(12,14)-prostaglandin J(2)
    Edward B Prage
    Departments of Chemistry and Biochemistry, Center in Molecular Toxicology, and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee 37232 0146, United States
    Biochemistry 51:2348-56. 2012
    ..The kinetics, regiochemistry, and stereochemistry of the spontaneous reaction of GSH with 15d-PGJ(2) were determined. The question of whether the anti-inflammatory properties of 15d-PGJ(2) are due to inhibition of MPGES1 is discussed...
  13. pmc Myristoylation exerts direct and allosteric effects on Gα conformation and dynamics in solution
    Anita M Preininger
    Vanderbilt University Medical Center, Nashville, Tennessee 37232, United States
    Biochemistry 51:1911-24. 2012
    ....
  14. pmc Location of inhibitor binding sites in the human inducible prostaglandin E synthase, MPGES1
    Edward B Prage
    Departments of Chemistry and Biochemistry, Center in Molecular Toxicology, and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee 37232 0146, United States
    Biochemistry 50:7684-93. 2011
    ....
  15. pmc Structure and function of YghU, a nu-class glutathione transferase related to YfcG from Escherichia coli
    Nina V Stourman
    Department of Biochemistry, Center in Molecular Toxicology, and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee 37232 0146, United States
    Biochemistry 50:1274-81. 2011
    ..The structures and properties of YghU and YfcG indicate that they are members of the same, but previously unidentified, subfamily of GSH transferase homologues, which we suggest be called the nu-class GSH transferases...
  16. pmc Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
    Johan Alander
    Institute of Environmental Medicine, Karolinska Institutet, SE 171 77 Stockholm, Sweden
    Arch Biochem Biophys 487:42-8. 2009
    ..5+/-0.5 mM). Thus we can reconcile previous observations and show here that MGST1 contains three active sites with different affinities for GSH and that only the high affinity site is catalytically competent...
  17. pmc Analysis of the structure and function of YfcG from Escherichia coli reveals an efficient and unique disulfide bond reductase
    Megan C Wadington
    Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232 0146, USA
    Biochemistry 48:6559-61. 2009
    ....