Mammalian class Sigma glutathione S-transferases: catalytic properties and tissue-specific expression of human and rat GSH-dependent prostaglandin D2 synthases

Ian R. Jowsey, Anne M. Thomson, Jack U. Flanagan, Paul R. Murdock, Gary B. T. Moore, David J. Meyer, Gregory J. Murphy, Stephen A. Smith, John D. Hayes

    Research output: Contribution to journalArticle

    82 Citations (Scopus)

    Abstract

    GSH-dependent prostaglandin D2 synthase (PGDS) enzymes represent the only vertebrate members of class Sigma glutathione S-transferases (GSTs) identified to date. Complementary DNA clones encoding the orthologous human and rat GSH-dependent PGDS (hPGDS and rPGDS, respectively) have been expressed in Escherichia coli, and the recombinant proteins isolated by affinity chromatography. The purified enzymes were both shown to catalyse specifically the isomerization of prostaglandin (PG) H2 to PGD2. Each transferase also exhibited GSH-conjugating and GSH-peroxidase activities. The ability of hPGDS to catalyse the conjugation of aryl halides and isothiocyanates with GSH was found to be less than that of the rat enzyme. Whilst there is no difference between the enzymes with respect to their Km values for 1-chloro-2,4-dinitrobenzene, marked differences were found to exist with respect to their Km for GSH (8mM versus 0.3mM for hPGDS and rPGDS, respectively). Using molecular modelling techniques, amino acid substitutions have been identified in the N-terminal domain of these enzymes that lie outside the proposed GSH-binding site, which may explain these catalytic differences. The tissue-specific expression of PGDS also varies significantly between human and rat; amongst the tissues examined, variation in expression between the two species was most apparent in spleen and bone marrow. Differences in catalytic properties and tissue-specific expression of hPGDS and rPGDS appears to reflect distinct physiological roles for class Sigma GST between species. The evolution of divergent functions for the hPGDS and rPGDS is discussed in the context of the orthologous enzyme from chicken.
    Original languageEnglish
    Pages (from-to)507-516
    Number of pages10
    JournalBiochemical Journal
    Volume359
    Issue number3
    DOIs
    Publication statusPublished - 1 Nov 2001

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    prostaglandin R2 D-isomerase
    Glutathione Transferase
    Rats
    Tissue
    Enzymes
    Prostaglandin H2
    Isothiocyanates
    Affinity chromatography
    Prostaglandin D2
    Dinitrochlorobenzene
    Molecular modeling
    Escherichia coli Proteins
    Amino Acid Substitution
    Transferases
    Isomerization
    Affinity Chromatography
    Recombinant Proteins
    Escherichia coli
    Peroxidase
    Vertebrates

    Cite this

    Jowsey, Ian R. ; Thomson, Anne M. ; Flanagan, Jack U. ; Murdock, Paul R. ; Moore, Gary B. T. ; Meyer, David J. ; Murphy, Gregory J. ; Smith, Stephen A. ; Hayes, John D. / Mammalian class Sigma glutathione S-transferases: catalytic properties and tissue-specific expression of human and rat GSH-dependent prostaglandin D2 synthases. In: Biochemical Journal. 2001 ; Vol. 359, No. 3. pp. 507-516.
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    title = "Mammalian class Sigma glutathione S-transferases: catalytic properties and tissue-specific expression of human and rat GSH-dependent prostaglandin D2 synthases",
    abstract = "GSH-dependent prostaglandin D2 synthase (PGDS) enzymes represent the only vertebrate members of class Sigma glutathione S-transferases (GSTs) identified to date. Complementary DNA clones encoding the orthologous human and rat GSH-dependent PGDS (hPGDS and rPGDS, respectively) have been expressed in Escherichia coli, and the recombinant proteins isolated by affinity chromatography. The purified enzymes were both shown to catalyse specifically the isomerization of prostaglandin (PG) H2 to PGD2. Each transferase also exhibited GSH-conjugating and GSH-peroxidase activities. The ability of hPGDS to catalyse the conjugation of aryl halides and isothiocyanates with GSH was found to be less than that of the rat enzyme. Whilst there is no difference between the enzymes with respect to their Km values for 1-chloro-2,4-dinitrobenzene, marked differences were found to exist with respect to their Km for GSH (8mM versus 0.3mM for hPGDS and rPGDS, respectively). Using molecular modelling techniques, amino acid substitutions have been identified in the N-terminal domain of these enzymes that lie outside the proposed GSH-binding site, which may explain these catalytic differences. The tissue-specific expression of PGDS also varies significantly between human and rat; amongst the tissues examined, variation in expression between the two species was most apparent in spleen and bone marrow. Differences in catalytic properties and tissue-specific expression of hPGDS and rPGDS appears to reflect distinct physiological roles for class Sigma GST between species. The evolution of divergent functions for the hPGDS and rPGDS is discussed in the context of the orthologous enzyme from chicken.",
    author = "Jowsey, {Ian R.} and Thomson, {Anne M.} and Flanagan, {Jack U.} and Murdock, {Paul R.} and Moore, {Gary B. T.} and Meyer, {David J.} and Murphy, {Gregory J.} and Smith, {Stephen A.} and Hayes, {John D.}",
    year = "2001",
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    Mammalian class Sigma glutathione S-transferases: catalytic properties and tissue-specific expression of human and rat GSH-dependent prostaglandin D2 synthases. / Jowsey, Ian R.; Thomson, Anne M.; Flanagan, Jack U.; Murdock, Paul R.; Moore, Gary B. T.; Meyer, David J. ; Murphy, Gregory J.; Smith, Stephen A.; Hayes, John D.

    In: Biochemical Journal, Vol. 359, No. 3, 01.11.2001, p. 507-516.

    Research output: Contribution to journalArticle

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    AU - Jowsey, Ian R.

    AU - Thomson, Anne M.

    AU - Flanagan, Jack U.

    AU - Murdock, Paul R.

    AU - Moore, Gary B. T.

    AU - Meyer, David J.

    AU - Murphy, Gregory J.

    AU - Smith, Stephen A.

    AU - Hayes, John D.

    PY - 2001/11/1

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    N2 - GSH-dependent prostaglandin D2 synthase (PGDS) enzymes represent the only vertebrate members of class Sigma glutathione S-transferases (GSTs) identified to date. Complementary DNA clones encoding the orthologous human and rat GSH-dependent PGDS (hPGDS and rPGDS, respectively) have been expressed in Escherichia coli, and the recombinant proteins isolated by affinity chromatography. The purified enzymes were both shown to catalyse specifically the isomerization of prostaglandin (PG) H2 to PGD2. Each transferase also exhibited GSH-conjugating and GSH-peroxidase activities. The ability of hPGDS to catalyse the conjugation of aryl halides and isothiocyanates with GSH was found to be less than that of the rat enzyme. Whilst there is no difference between the enzymes with respect to their Km values for 1-chloro-2,4-dinitrobenzene, marked differences were found to exist with respect to their Km for GSH (8mM versus 0.3mM for hPGDS and rPGDS, respectively). Using molecular modelling techniques, amino acid substitutions have been identified in the N-terminal domain of these enzymes that lie outside the proposed GSH-binding site, which may explain these catalytic differences. The tissue-specific expression of PGDS also varies significantly between human and rat; amongst the tissues examined, variation in expression between the two species was most apparent in spleen and bone marrow. Differences in catalytic properties and tissue-specific expression of hPGDS and rPGDS appears to reflect distinct physiological roles for class Sigma GST between species. The evolution of divergent functions for the hPGDS and rPGDS is discussed in the context of the orthologous enzyme from chicken.

    AB - GSH-dependent prostaglandin D2 synthase (PGDS) enzymes represent the only vertebrate members of class Sigma glutathione S-transferases (GSTs) identified to date. Complementary DNA clones encoding the orthologous human and rat GSH-dependent PGDS (hPGDS and rPGDS, respectively) have been expressed in Escherichia coli, and the recombinant proteins isolated by affinity chromatography. The purified enzymes were both shown to catalyse specifically the isomerization of prostaglandin (PG) H2 to PGD2. Each transferase also exhibited GSH-conjugating and GSH-peroxidase activities. The ability of hPGDS to catalyse the conjugation of aryl halides and isothiocyanates with GSH was found to be less than that of the rat enzyme. Whilst there is no difference between the enzymes with respect to their Km values for 1-chloro-2,4-dinitrobenzene, marked differences were found to exist with respect to their Km for GSH (8mM versus 0.3mM for hPGDS and rPGDS, respectively). Using molecular modelling techniques, amino acid substitutions have been identified in the N-terminal domain of these enzymes that lie outside the proposed GSH-binding site, which may explain these catalytic differences. The tissue-specific expression of PGDS also varies significantly between human and rat; amongst the tissues examined, variation in expression between the two species was most apparent in spleen and bone marrow. Differences in catalytic properties and tissue-specific expression of hPGDS and rPGDS appears to reflect distinct physiological roles for class Sigma GST between species. The evolution of divergent functions for the hPGDS and rPGDS is discussed in the context of the orthologous enzyme from chicken.

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    M3 - Article

    VL - 359

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    EP - 516

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