Home LiteratureArticle Details
PMID: 9307035 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Evidence that human class Theta glutathione S-transferase T1-1 can catalyse the activation of dichloromethane, a liver and lung carcinogen in the mouse. Comparison of the tissue distribution of GST T1-1 with that of classes Alpha, Mu and Pi GST in human.

The Biochemical journal ·Vol. 326 ( Pt 3) ·1997-09-15 ·Pages 837-46

Sherratt PJ, Pulford DJ, Harrison DJ, Green T, Hayes JD

Abstract

The cDNA encoding human glutathione S-transferase (GST) T1 has been expressed as two recombinant forms in Escherichia coli that could be purified by affinity chromatography on either IgG-Sepharose or nickel-agarose; one form of the transferase was synthesized from the pALP 1 expression vector as a Staphylococcus aureus protein A fusion, whereas the other form was synthesized from the pET-20b expression vector as a C-terminal polyhistidine-tagged recombinant. The yields of the two purified recombinant proteins from E. coli cultures were approx. 15 mg/l for the protein A fusion and 25 mg/l for the C-terminal polyhistidine-tagged GST T1-1. The purified recombinant proteins were catalytically active, although the protein A fusion was typically only 5-30% as active as the histidine-tagged GST. Both recombinant forms could catalyse the conjugation of glutathione with the model substrates 1,2-epoxy-3-(4'-nitrophenoxy)propane,4-nitrobenzyl chloride and 4-nitrophenethyl bromide but were inactive towards 1-chloro-2,4-dinitrobenzene, ethacrynic acid and 1-menaphthyl sulphate. Recombinant human GST T1-1 was found to exhibit glutathione peroxidase activity and could catalyse the reduction of cumene hydroperoxide. In addition, recombinant human GST T1-1 was found to conjugate glutathione with dichloromethane, a pulmonary and hepatic carcinogen in the mouse. Immunoblotting with antibodies raised against different transferase isoenzymes showed that GST T1-1 is expressed in a large number of human organs in a tissue-specific fashion that differs from the pattern of expression of classes Alpha, Mu and Pi GST. Most significantly, GST T1-1 was found in only low levels in human pulmonary soluble extract of cells, suggesting that in man the lung has little capacity to activate the volatile dichloromethane.

MeSH Terms
Amino Acid Sequence Animals Carcinogens/metabolism,pharmacology Cloning, Molecular Glutathione Transferase/metabolism Humans Liver/drug effects,pathology Lung/drug effects,pathology Methylene Chloride/metabolism,pharmacology Mice Molecular Sequence Data Organ Specificity Recombinant Proteins/genetics,metabolism
Chemicals
Carcinogens Recombinant Proteins Methylene Chloride Glutathione Transferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sherratt P J
Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, Scotland, U.K.
Pulford D J
Harrison D J
Green T
Hayes J D
References (56)
56 references, click to expand
  1. Purification and physical characterization of glutathione S-transferase K. Differential use of S-hexylglutathione and glutathione affinity matrices to isolate a novel glutathione S-transferase from rat liver.
    Biochem J. 1986 Feb 1;233(3):789-98 PMID: 3707526
  2. Use of immuno-blot techniques to discriminate between the glutathione S-transferase Yf, Yk, Ya, Yn/Yb and Yc subunits and to study their distribution in extrahepatic tissues. Evidence for three immunochemically distinct groups of transferase in the rat.
    Biochem J. 1986 Feb 1;233(3):779-88 PMID: 3707525
  3. Isozyme specificity of rat liver glutathione S-transferases in the formation of PGF2 alpha and PGE2 from PGH2.
    Arch Biochem Biophys. 1987 Dec;259(2):548-57 PMID: 3480701
  4. Amino acid substitutions in the human glutathione S-transferases confer different specificities in the prostaglandin endoperoxide conversion pathway.
    Biochem Biophys Res Commun. 1989 Jan 31;158(2):497-502 PMID: 2916997
  5. Single-step purification and h.p.l.c. analysis of glutathione transferase 8-8 in rat tissues.
    Biochem J. 1989 Jun 15;260(3):785-8 PMID: 2764904
  6. Glutathione S-transferase and glutathione peroxidase expression in normal and tumour human tissues.
    Carcinogenesis. 1990 Mar;11(3):451-8 PMID: 2311189
  7. A new class of rat glutathione S-transferase Yrs-Yrs inactivating reactive sulfate esters as metabolites of carcinogenic arylmethanols.
    J Biol Chem. 1990 Jul 15;265(20):11973-81 PMID: 2114406
  8. Variation in the expression of Mu-class glutathione S-transferase isoenzymes from human skeletal muscle. Evidence for the existence of heterodimers.
    Biochem J. 1991 Jan 15;273(Pt 2):323-32 PMID: 1991033
  9. Theta, a new class of glutathione transferases purified from rat and man.
    Biochem J. 1991 Mar 1;274 ( Pt 2):409-14 PMID: 1848757
  10. Role of cysteine residues in the activity of rat glutathione transferase P (7-7): elucidation by oligonucleotide site-directed mutagenesis.
    Biochem Biophys Res Commun. 1991 Sep 16;179(2):790-7 PMID: 1898401
  11. Protein A-calmodulin fusions: a novel approach for investigating calmodulin function in yeast.
    Mol Microbiol. 1992 Mar;6(6):703-13 PMID: 1573999
  12. Dissociation of a new glutathione S-transferase activity in human erythrocytes.
    Biochem Pharmacol. 1992 Apr 15;43(8):1671-4 PMID: 1575765
  13. Expression of mammalian glutathione S-transferase 5-5 in Salmonella typhimurium TA1535 leads to base-pair mutations upon exposure to dihalomethanes.
    Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8576-80 PMID: 8378332
  14. Human Mu-class glutathione S-transferases present in liver, skeletal muscle and testicular tissue.
    Biochim Biophys Acta. 1993 Nov 10;1203(1):131-41 PMID: 8218382
  15. Significance of an unusually low Km for glutathione in glutathione transferases of the alpha, mu and pi classes.
    Xenobiotica. 1993 Aug;23(8):823-34 PMID: 8284939
  16. Detoxication of base propenals and other alpha, beta-unsaturated aldehyde products of radical reactions and lipid peroxidation by human glutathione transferases.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1480-4 PMID: 8108434
  17. A novel glutathione S-transferase isozyme similar to GST 8-8 of rat and mGSTA4-4 (GST 5.7) of mouse is selectively expressed in human tissues.
    Biochim Biophys Acta. 1994 Feb 16;1204(2):279-86 PMID: 8142470
  18. Human glutathione S-transferase theta (GSTT1): cDNA cloning and the characterization of a genetic polymorphism.
    Biochem J. 1994 May 15;300 ( Pt 1):271-6 PMID: 8198545
  19. Characterization of a human class-Theta glutathione S-transferase with activity towards 1-menaphthyl sulphate.
    Biochem J. 1992 Sep 15;286 ( Pt 3):929-35 PMID: 1417752
  20. Relationship between hepatic DNA damage and methylene chloride-induced hepatocarcinogenicity in B6C3F1 mice.
    Carcinogenesis. 1994 May;15(5):991-6 PMID: 8200106
  21. Glutathione-associated enzymes in anticancer drug resistance.
    Cancer Res. 1994 Aug 15;54(16):4313-20 PMID: 8044778
  22. Expression cloning of a cDNA for human leukotriene C4 synthase, an integral membrane protein conjugating reduced glutathione to leukotriene A4.
    Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7663-7 PMID: 8052639
  23. Enzymology of microsomal glutathione S-transferase.
    Adv Pharmacol. 1994;27:19-35 PMID: 8068553
  24. Molecular cloning and expression of human leukotriene-C4 synthase.
    Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9745-9 PMID: 7937884
  25. Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport.
    Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):13033-7 PMID: 7809167
  26. Crystal structure of a theta-class glutathione transferase.
    EMBO J. 1995 May 15;14(10):2133-43 PMID: 7774571
  27. Gene deletion of glutathione S-transferase theta: correlation with induced genetic damage and potential role in endogenous mutagenesis.
    Cancer Epidemiol Biomarkers Prev. 1995 Apr-May;4(3):253-9 PMID: 7606200
  28. Methylene chloride-induced DNA damage: an interspecies comparison.
    Carcinogenesis. 1995 Aug;16(8):1919-26 PMID: 7634422
  29. Oxidation of 1,2-epoxy-3-butene to 1,2:3,4-diepoxybutane by cDNA-expressed human cytochromes P450 2E1 and 3A4 and human, mouse and rat liver microsomes.
    Carcinogenesis. 1995 Oct;16(10):2287-93 PMID: 7586124
  30. Characterization of rat spleen prostaglandin H D-isomerase as a sigma-class GSH transferase.
    Biochem J. 1995 Nov 1;311 ( Pt 3):739-42 PMID: 7487927
  31. Enhancement of bacterial mutagenicity of bifunctional alkylating agents by expression of mammalian glutathione S-transferase.
    Chem Res Toxicol. 1995 Apr-May;8(3):465-72 PMID: 7578934
  32. Human glutathione S-transferase T1-1 enhances mutagenicity of 1,2-dibromoethane, dibromomethane and 1,2,3,4-diepoxybutane in Salmonella typhimurium.
    Carcinogenesis. 1996 Jan;17(1):163-6 PMID: 8565128
  33. Heterologous expression, purification and characterization of rat class theta glutathione transferase T2-2.
    Biochem J. 1996 May 15;316 ( Pt 1):131-6 PMID: 8645195
  34. Production and characterization of monoclonal antibodies against class theta glutathione S-transferase T1-1.
    Hybridoma. 1996 Feb;15(1):77-82 PMID: 9064289
  35. Isolation of a mouse theta glutathione S-transferase active with methylene chloride.
    Biochem J. 1996 Mar 1;314 ( Pt 2):445-8 PMID: 8670055
  36. Regulation of carbonyl-reducing enzymes in rat liver by chemoprotectors.
    Cancer Res. 1996 Jun 15;56(12):2758-66 PMID: 8665510
  37. Identification and characterization of a novel human microsomal glutathione S-transferase with leukotriene C4 synthase activity and significant sequence identity to 5-lipoxygenase-activating protein and leukotriene C4 synthase.
    J Biol Chem. 1996 Sep 6;271(36):22203-10 PMID: 8703034
  38. The distribution of theta-class glutathione S-transferases in the liver and lung of mouse, rat and human.
    Biochem J. 1996 Aug 15;318 ( Pt 1):297-303 PMID: 8761485
  39. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance.
    Crit Rev Biochem Mol Biol. 1995;30(6):445-600 PMID: 8770536
  40. Purification, characterization and tissue distribution of human class theta glutathione S-transferase T1-1.
    Biochem Mol Biol Int. 1996 May;39(1):21-9 PMID: 8799324
  41. Glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) genetic polymorphism and susceptibility to gastric and colorectal adenocarcinoma.
    Carcinogenesis. 1996 Sep;17(9):1855-9 PMID: 8824506
  42. Multiple cutaneous basal cell carcinomas: glutathione S-transferase (GSTM1, GSTT1) and cytochrome P450 (CYP2D6, CYP1A1) polymorphisms influence tumour numbers and accrual.
    Carcinogenesis. 1996 Sep;17(9):1891-6 PMID: 8824510
  43. Purification and characterization of a new glutathione S-transferase, class theta, from human erythrocytes.
    Arch Toxicol. 1996;70(9):559-66 PMID: 8831906
  44. Mutagenesis of the active site of the human Theta-class glutathione transferase GSTT2-2: catalysis with different substrates involves different residues.
    Biochem J. 1996 Oct 1;319 ( Pt 1):315-21 PMID: 8870684
  45. Glutathione S-transferase class Kappa: characterization by the cloning of rat mitochondrial GST and identification of a human homologue.
    Biochem J. 1996 Nov 1;319 ( Pt 3):749-54 PMID: 8920976
  46. Subunit diversity and tissue distribution of human glutathione S-transferases: interpretations based on electrospray ionization-MS and peptide sequence-specific antisera.
    Biochem J. 1997 Jul 15;325 ( Pt 2):481-6 PMID: 9230131
  47. The colorimetric estimation of formaldehyde by means of the Hantzsch reaction.
    Biochem J. 1953 Oct;55(3):416-21 PMID: 13105648
  48. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  49. Enzymatic conjugation of epoxides with glutathione.
    J Biol Chem. 1973 May 25;248(10):3702-7 PMID: 4634012
  50. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  51. Assays for differentiation of glutathione S-transferases.
    Methods Enzymol. 1981;77:398-405 PMID: 7329316
  52. Carcinogenicity and toxicity of 1,2-dibromoethane in the rat.
    Toxicol Appl Pharmacol. 1982 Apr;63(2):155-65 PMID: 7046137
  53. Purification and characterization of three forms of glutathione S-transferase A. A comparative study of the major YaYa-, YbYb- and YcYc-containing glutathione S-transferases.
    Biochem J. 1982 Dec 1;207(3):459-70 PMID: 7165703
  54. Purification of human hepatic glutathione S-transferases and the development of a radioimmunoassay for their measurement in plasma.
    Clin Chim Acta. 1983 Oct 31;134(1-2):107-21 PMID: 6652905
  55. Methylene chloride: a two-year inhalation toxicity and oncogenicity study in rats and hamsters.
    Fundam Appl Toxicol. 1984 Feb;4(1):30-47 PMID: 6693002
  56. Physiologically based pharmacokinetics and the risk assessment process for methylene chloride.
    Toxicol Appl Pharmacol. 1987 Feb;87(2):185-205 PMID: 3824380
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3260837
Published
1997-09-15
Pages
837-46
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218740
Subset
IM
External Links
PubMed source DOI: 10.1042/bj3260837 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2