Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.5.1.18 (glutathione S-transferase)
22,582 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

5-Lipoxygenase-activating protein (FLAP) and leukotriene C4 (LTC4) synthase, two proteins involved in leukotriene biosynthesis, have been demonstrated to be 31% identical at the amino acid level. We have recently identified and characterized a novel member of the FLAP/LTC4 synthase gene family termed microsomal glutathione S-transferase II (microsomal GST-II). The open reading frame encodes a 16.6-kDa protein with a calculated pI of 10.4. Microsomal GST-II has 33% amino acid identity to FLAP, 44% amino acid identity to LTC4 synthase, and 11% amino acid identity to the previously characterized human microsomal GST (microsomal GST-I). Microsomal GST-II also has a similar hydrophobicity pattern to FLAP, LTC4 synthase, and microsomal GST-I. Fluorescent in situ hybridization mapped microsomal GST-II to chromosomal localization 4q28-31. Microsomal GST-II has a wide tissue distribution (at the mRNA level) and was specifically expressed in human liver, spleen, skeletal muscle, heart, adrenals, pancreas, prostate, testis, fetal liver, and fetal spleen. In contrast, microsomal GST-II mRNA expression was very low (when present) in lung, brain, placenta, and bone marrow. This differs from FLAP mRNA, which was detected in lung, various organs of the immune system, and peripheral blood leukocytes, and LTC4 synthase mRNA, which could not be detected in any tissues by Northern blot analysis. Microsomal GST-II and LTC4 synthase were expressed in a baculovirus insect cell system, and microsomes from Sf9 cells containing microsomal GST-II or LTC4 synthase were both found to catalyze the production of LTC4 from LTA4 and reduced glutathione. Microsomal GST-II also catalyzed the formation of another product, displaying a conjugated triene UV absorption spectra with a maximum at 283 nm, suggesting less catalytic stereospecificity compared with LTC4 synthase. Also, the apparent Km for LTA4 was higher for microsomal GST-II (41 microM) than LTC4 synthase (7 microM). In addition, unlike LTC4 synthase, microsomal GST-II was able to catalyze the conjugation of 1-chloro-2, 4-dinitrobenzene with reduced glutathione. Therefore, it is proposed that this novel membrane protein is a member of the microsomal glutathione S-transferase family, also including LTC4 synthase, with significant sequence identities to both LTC4 synthase and FLAP.
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PMID: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. 870 34

Microsomal glutathione S-transferase-II (GST-II) has recently been discovered and characterized as a member of the 5-lipoxygenase-activating protein (FLAP)/5(S)-hydroxy-6(R)-S-glutathionyl-7,9-trans-11, 14-cis-eicosatetraenoic acid (LTC4) synthase gene family, which also includes microsomal glutathione S-transferase-I (GST-I) as a distant member of this gene family. This new enzyme is unique as it is the only member of this family capable of efficiently conjugating reduced glutathione to both 5,6-oxido-7,9,11,14-eicosatetraenoic acid (LTA4) and 1-chloro-2,4-dinitrobenzene. Although microsomal GST-II has been demonstrated to display both general glutathione S-transferase (GST) and specific LTC4 synthase activities, its biological function remains unknown. In this study, we investigated the physiological location of microsomal GST-II as well as the relative importance of this enzyme versus LTC4 synthase for the production of LTC4 in various human tissues and cells that have been previously demonstrated to possess LTC4 synthase activity. As determined by Western blot, microsomal GST-II was predominantly expressed in human liver microsomes, human endothelial cell membranes, and sparsely detected in human lung membranes. In contrast, LTC4 synthase was prevalent in human lung membranes, human platelet homogenates, and human kidney tissue. Concomitant to the formation of LTC4, microsomal GST-II also produces a new metabolite of LTA4, a postulated LTC4 isomer. This isomer was used to distinguish between microsomal GST-II and LTC4 synthase activities involved in the biosynthesis of LTC4. Based on the relative production of LTC4 to the LTC4 isomer, microsomal GST-II was demonstrated to be the principal enzyme responsible for LTC4 production in human liver microsomes and human endothelial cells and played a minor role in the formation of LTC4 in human lung membranes. In comparison, LTC4 synthase was the main enzyme capable of catalyzing the conjugation of reduced glutathione to LTA4 in human lung membranes and human platelet homogenates. Therefore, microsomal GST-II appears to be an integral component in the detoxification of biological systems due to its marked presence in human liver, in accordance with its known GST activity. Microsomal GST-II, however, may also be pivotal for cysteinyl leukotriene formation in endothelial cells, and this could change our current understanding of the regulation of leukotriene biosynthesis in inflammatory disorders such as asthma.
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PMID:Production of leukotriene C4 in different human tissues is attributable to distinct membrane bound biosynthetic enzymes. 909 65

Leukotriene (LT) C4 synthase catalyzes the conjugation of LTA4 with reduced glutathione (GSH) to form LTC4, the parent compound of cysteinyl leukotrienes. It is a 18 kDa protein that functions as homodimer. Cloning of LTC4 synthase cDNA reveals amino acid homology with 5-lipoxygenase activating protein (FLAP) and newly identified microsomal glutathione S-transferase II (mGST-II) but not with cytosolic GSTs or mGST-I. LTC4 synthase gene contains 5 exons and four introns. This gene has been localized to the long arm of human chromosome 5 at the region of 5q35 which is in close proximity to the cluster of genes that are involved in inflammation and asthma. Mutagenic studies reveals that amino acid residues Arg-51 and Tyr-93 are critical for catalytic function. Arg-51 was proposed to open the epoxide ring of LTA4 and Tyr-93 to provide the thiolate anion of GSH.
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PMID:Leukotriene C4 synthase: a critical enzyme for the biosynthesis of SRS-A. 923 64

5-Lipoxygenase activating protein (FLAP), leukotriene-C4 (LTC4) synthase, and microsomal glutathione S-transferase II (microsomal GST-II) are all members of a common gene family that may also include microsomal GST-I. The present work describes the identification and characterization of a novel member of this family termed microsomal glutathione S-transferase III (microsomal GST-III). The open reading frame encodes a 16.5-kDa protein with a calculated pI of 10.2. Microsomal GST-III has 36, 27, 22, and 20% amino acid identity to microsomal GST-II, LTC4 synthase, microsomal GST-I, and FLAP, respectively. Microsomal GST-III also has a similar hydrophobicity pattern to FLAP, LTC4 synthase, and microsomal GST-I. Fluorescent in situ hybridization mapped microsomal GST-III to chromosomal localization 1q23. Like microsomal GST-II, microsomal GST-III has a wide tissue distribution (at the mRNA level) and is predominantly expressed in human heart, skeletal muscle, and adrenal cortex, and it is also found in brain, placenta, liver, and kidney tissues. Expression of microsomal GST-III mRNA was also detected in several glandular tissues such as pancreas, thyroid, testis, and ovary. In contrast, microsomal GST-III mRNA expression was very low (if any) in lung, thymus, and peripheral blood leukocytes. Microsomal GST-III protein was expressed in a baculovirus insect cell system, and microsomes from Sf9 cells containing either microsomal GST-II or microsomal GST-III were both found to possess glutathione-dependent peroxidase activity as shown by their ability to reduce 5-HPETE to 5-HETE in the presence of reduced glutathione. The apparent Km of 5-HPETE was determined to be approximately 7 microM for microsomal GST-II and 21 microM for microsomal GST-III. Microsomal GST-III was also found to catalyze the production of LTC4 from LTA4 and reduced glutathione. Based on these catalytic activities it is proposed that this novel membrane protein is a member of the microsomal glutathione S-transferase super family, which also includes microsomal GST-I, LTC4 synthase, FLAP, and microsomal GST-II.
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PMID:Identification and characterization of a novel microsomal enzyme with glutathione-dependent transferase and peroxidase activities. 927 57

Leukotriene C4 (LTC4) synthase (LTC4S), an integral membrane protein, catalyzes the conjugation of leukotriene A4 with reduced glutathione to form LTC4, the biosynthetic parent of the additional cysteinyl leukotriene metabolites. An XmnI-digested fragment of a P1 clone from a 129 mouse ES library contained the full-length gene of 2.01 kb for mouse LTC4S. The mouse LTC4S gene is comprised of 5 exons of 122, 100, 71, 82 and 241 nucleotides, with intron sizes that range from 76 nucleotides to 937 nucleotides. The intron/exon boundaries are identical to those of the human genes for LTC4S and 5-lipoxygenase-activating protein (FLAP). Primer extension demonstrated a single transcription-initiation site 64 bp 5' of the ATG translation-start site. Nucleotide sequencing of 1.2 kb of the 5' flanking region revealed multiple putative sites for activating protein-2, CCAAT/enhancer-binding protein, and polyoma virus enhancer-3. Fluorescent in situ hybridization mapped the mouse LTC4S gene to mouse chromosome 11, in a region containing the genes for interleukin 13 and granulocyte/macrophage-colony-stimulating factor, and orthologous to the chromosomal location of 5q35 for the human LTC4S gene. Thus, the mouse LTC4S gene is similar in size, intron/exon organization and chromosomal localization to the human LTC4S gene. Recent mutagenic analysis of the conjugation function of human LTC4S has identified R51 and Y93 as critical for acid and base catalysis of LTA4 and reduced glutathione, respectively. A comparison across species for proteins that possess LTC4S activity reveals conservation of both of these residues, whereas R51 is absent in the FLAP molecules. Thus, within the glutathione S-transferase superfamily of genes, alignment of specific residues allows the separation of LTC4S family members from their most structurally similar counterparts, the FLAP molecules.
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PMID:Molecular cloning of the gene for mouse leukotriene-C4 synthase. 934 32

gamma-Carboxylation of vitamin K-dependent proteins requires a functional vitamin K cycle to produce the active vitamin K cofactor for the gamma-carboxylase which posttranslationally modifies precursors of these proteins to contain gamma-carboxyglutamic acid residues. The warfarin-sensitive enzyme vitamin K epoxide reductase (VKOR) of the cycle reduces vitamin K 2,3-epoxide to the active vitamin K hydroquinone cofactor. Because of the importance of warfarin as an anticoagulant in prophylactic medicine and as a poison in rodent pest control, numerous attempts have been made to understand the molecular mechanism underlying warfarin-sensitive vitamin K 2,3-epoxide reduction. In search for protein components that could be involved in this reaction we designed an in vitro gamma-carboxylation test system where the warfarin-sensitive VKOR produces the cofactor for the gamma-carboxylase. Dissection of this system by chromatographic techniques has identified a member(s) of the glutathione S-transferase gene family as one component of the VKOR enzyme complex in the endoplasmic reticulum membrane. The affinity-purified glutathione S-transferase(s) was sensitive to warfarin but lost its warfarin sensitivity and glutathione S-transferase activity upon association with lipids in the presence of Mn2+ or Ca2+. In the gamma-carboxylation test system, loss of warfarin-sensitive glutathione S-transferase activity coincided with formation of the VKOR enzyme complex. It is proposed that formation of VKOR in the endoplasmic reticulum membrane resembles formation of the lipoxygenase enzyme complex where the glutathione S-transferase-related FLAP protein binds cytosolic lipoxygenase to form a membrane enzyme complex.
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PMID:Assembly of the warfarin-sensitive vitamin K 2,3-epoxide reductase enzyme complex in the endoplasmic reticulum membrane. 936 Sep 81

Microsomal glutathione transferase-1 (MGST-1) is an abundant protein that catalyzes the conjugation of electrophilic compounds with glutathione, as well as the reduction of lipid hydroperoxides. Here we report that leukotriene C4 is a potent inhibitor of MGST-1. Leukotriene C4 was found to be a tight-binding inhibitor, with a Ki of 5.4 nM for the unactivated enzyme, and 9.2 nM for the N-ethylmaleimide activated enzyme. This is the first tight-binding inhibitor characterized for this enzyme. Leukotriene C4 was competitive with respect to glutathione and non-competitive toward the second substrate, CDNB. Analysis of stoichiometry supports binding of one molecule of inhibitor per homotrimer. Leukotrienes A4, D4, and E4 were much weaker inhibitors of the purified enzyme (by at least 3 orders of magnitude). Leukotriene C4 analogues, which have been developed as antagonists of leukotriene receptors, were found to display varying degrees of inhibition of MGST-1. In particular, the cysteinyl-leukotriene analogues SKF 104,353, ONO-1078, and BAYu9773 were strong inhibitors (IC50 values: 0.13, 3. 7, and 7.6 microM, respectively). In view of the partial structural similarity between MGST-1, leukotriene C4 synthase, and 5-lipoxygenase activating protein (FLAP), it was of interest that leukotriene C4 synthesis inhibitors (which antagonize FLAP) also displayed significant inhibition (e.g. IC50 for BAYx1005 was 58 microM). In contrast, selective 5-lipoxygenase inhibitors such as zileuton only marginally inhibited activity at high concentrations (500 microM). Our discovery that leukotriene C4 and drugs developed based on its structure are potent inhibitors of MGST-1 raises the possibility that MGST-1 influences the cellular processing of leukotrienes. These findings may also have implications for the effects and side-effects of drugs developed to manipulate leukotrienes.
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PMID:Leukotriene C4 is a tight-binding inhibitor of microsomal glutathione transferase-1. Effects of leukotriene pathway modifiers. 989 Sep 56

LTC4S conjugates reduce glutathione to LTA4 and is positioned as the pivotal and only committed enzyme involved in the formation of cysteinyl LTs. Despite its function as an enzyme that conjugates glutathione to LTA4, it is abundantly clear that LTC4S differs from the classic glutathione S-transferase (GST) families. This distinction is based on narrow substrate specificity, inability to conjugate GSH to xenobiotics, differential susceptibility to inhibitors, lack of homology, and failure to be immunorecognized by specific microsomal GST antibodies. The presence of LTC4S protein is restricted to a limited number of hematopoietic cells to include mast cells, eosinophils, basophils, monocytes/macrophages, and platelets, with the platelet being unique in its lack of the complete biosynthetic pathway for cysteinyl LTs. The purification of the protein and the cloning of the cDNA have demonstrated that the kinetic parameters of LTC4S are similar for the isolated natural or recombinant proteins. The protein is an 18-kDa integral perinuclear membrane enzyme, which is functional as a homodimer. The cDNA encodes a 150 amino-acid polypeptide monomer with three hydrophobic domains interspersed by two hydrophilic loops. Homology and secondary structural predictions have revealed that LTC4S is a member of a novel gene family that includes FLAP, mGST II, and mGST III. Each of these molecules is an integral membrane protein with the capacity to participate in LT biosynthesis: LTC4S as the terminal and only committed enzyme in cysteinyl LT formation, FLAP as an arachidonic acid presentation protein, and mGST II and mGST III as unique dual-function enzymes with primary detoxification functions. Site directed mutagenic studies of LTC4S have revealed that two residues, R51 and Y93, are involved in the acid and base catalysis, respectively, of LTA4 and GSH. Alignment of molecules with LTA4 conjugating ability demonstrates conservation of amino acid residues R51 and Y93, which appear necessary for this specific enzymatic function. The 2.5-Kb gene for human LTC4S contains five small exons and four introns, and the 5' UTR contains consensus sequences for AP-1 and AP-2 sites as well as an SP-1 site. The chromosomal localization of this gene is 5q35, distal to that of cytokine, growth factor, and receptor genes that have relevance to the development of allergic inflammation. Furthermore, there is genetic linkage of this region of human chromosome 5 to atopy and asthma, whereas no linkage exists for the chromosomal localization of the other family members, FLAP and mGST II, distinguishing LTC4S as a unique member of the novel gene family. LTC4S is profoundly overexpressed in the aspirin-induced asthmatic phenotype and correlates with overproduction of cysteinyl LTs and bronchial hyperreactivity to lysine aspirin. Ongoing studies are directed to the genomic regulation and additional polymorphisms within the gene of this pivotal enzyme, as well as to further identification of the amino acid residues central to its catalytic function.
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PMID:LTC4 synthase. Enzymology, biochemistry, and molecular characterization. 1043 63

In this paper we report the genomic organization of the human microsomal GST-I gene. This gene spans 18 kb, and contains seven exons. Sequences that encode the 155 amino acid open reading frame are present in Exons II, III, IV, the 5'-untranslated region is present in Exons Ia, Ib, Ic, Id, and II, and the 3'-untranslated region is present in Exon IV. Exons Ia, Ib, Ic, Id, and III are alternatively spliced to generate at least six different mGST-I transcripts. The results of EST and PCR analysis show that most mGST-I transcripts terminate within Exon Ib, and primer extension analysis shows these transcripts initiate at three major sites located at 79, 81, and 88 nucleotides upstream of the ATG initiation codon. Sequences surrounding the putative initiation sites are G-C rich, and several Sp1 consensus binding sites were identified. Northern analysis shows that the human GST-I gene is preferentially expressed as a 1.0 kb transcript in liver, and in several other tissues. Finally, a comparison of the mGST-I and PIG12 sequences with those of FLAP, LTC4 synthase, mGST-II, and mGST-III suggests that these proteins are the related products of a dispersed microsomal GST gene superfamily.
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PMID:Microsomal GST-I: genomic organization, expression, and alternative splicing of the human gene. 1052 15

1. We investigated the role of arachidonic acid metabolism and assessed the participation of mast cells and leukocytes in neurogenic inflammation in rat paw skin. We compared the effect of lipoxygenase (LOX) and cyclo-oxygenase (COX) inhibitors on oedema induced by saphenous nerve stimulation, substance P (SP), and compound 48/80. 2. Intravenous (i.v.) pre-treatment with a dual COX/LOX inhibitor (RWJ 63556), a dual LOX inhibitor/cysteinyl-leukotriene (CysLt) receptor antagonist (Rev 5901), a LOX inhibitor (AA 861), a five-lipoxygenase activating factor (FLAP) inhibitor (MK 886), or a glutathione S-transferase inhibitor (ethacrynic acid) significantly inhibited (40 to 60%) the development of neurogenic oedema, but did not affect cutaneous blood flow. Intradermal (i.d.) injection of LOX inhibitors reduced SP-induced oedema (up to 50% for RWJ 63556 and MK 886), whereas ethacrynic acid had a potentiating effect. 3. Indomethacin and rofecoxib, a highly selective COX-2 inhibitor, did not affect neurogenic and SP-induced oedema. Surprisingly, the structurally related COX-2 inhibitors, NS 398 and nimesulide, significantly reduced both neurogenic and SP-induced oedema (70% and 42% for neurogenic oedema, respectively; 49% and 46% for SP-induced oedema, respectively). 4. COX-2 mRNA was undetectable in saphenous nerves and paw skin biopsy samples, before and after saphenous nerve stimulation. 5. A mast cell stabilizer, cromolyn, and a H(1) receptor antagonist, mepyramine, significantly inhibited neurogenic (51% and 43%, respectively) and SP-induced oedema (67% and 63%, respectively). 6. The co-injection of LOX inhibitors and compound 48/80 did not alter the effects of compound 48/80. Conversely, ethacrynic acid had a significant potentiating effect. The pharmacological profile of the effect of COX inhibitors on compound 48/80-induced oedema was similar to that of neurogenic and SP-induced oedema. 7. The polysaccharide, fucoidan (an inhibitor of leukocyte rolling) did not affect neurogenic or SP-induced oedema. 8. Thus, (i) SP-induced leukotriene synthesis is involved in the development of neurogenic oedema in rat paw skin; (ii) this leukotriene-mediated plasma extravasation might be independent of mast cell activation and/or of the adhesion of leukocytes to the endothelium; (iii) COX did not appear to play a significant role in this process.
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PMID:Cyclo-oxygenase and lipoxygenase pathways in mast cell dependent-neurogenic inflammation induced by electrical stimulation of the rat saphenous nerve. 1126 53


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