Structures of class pi glutathione S-transferase from human placenta in complex with substrate, transition-state analogue and inhibitor

Prade, Lars; Huber, Robert; Th, Manoharan; We, Fahl; Reuter, W

doi:10.1016/s0969-2126(97)00281-5

Cited by 76 publications

(64 citation statements)

References 35 publications

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“…Tyr 152 , together with Arg 14 , Glu 96 , and two water molecules bound at the bottom of the catalytic cleft (Fig. 1C), is thought to form the hydrogen bonding network involved in the binding of GSH, as reported in the Pi class GSTs (40). The partial decrease in both PGDS and GST activities of Y152F (Fig.…”

mentioning

confidence: 53%

Structural Basis of Hematopoietic Prostaglandin D Synthase Activity Elucidated by Site-directed Mutagenesis

Pinzar¹,

Miyano²,

Kanaoka³

et al. 2000

Journal of Biological Chemistry

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Hematopoietic prostaglandin (PG) D synthase (PGDS)is the first identified vertebrate ortholog in the Sigma class of the glutathione S-transferase (GST) family and catalyzes both isomerization of PGH 2 to PGD 2 and conjugation of glutathione to 1-chloro-2,4-dinitrobenzene. We introduced site-directed mutations of 199, which are presumed to participate in catalysis or PGH 2 substrate binding based on the crystallographic structure. Mutants were analyzed in terms of structure, GST and PGDS activities, and activation of the glutathione thiol group. Of all the mutants, only Y8F, W104I, K112E, and L199F showed minor but substantial differences in their far-UV circular dichroism spectra from the wild-type enzyme. Y8F, R14K/E, and W104I were completely inactive. C156L/Y selectively lost only PGDS activity. K112E reduced GST activity slightly and PGDS activity markedly, whereas K198E caused a selective decrease in PGDS activity and K m for glutathione and PGH 2 in the PGDS reaction. No significant changes were observed in the catalytic activities of Y152F and L199F, although their K m for glutathione was increased. Using 5,5-dithiobis(2-nitrobenzoic acid) as an SH-selective agent, we found that only Y8F and R14E/K did not accelerate the reactivity of the glutathione thiol group under the low reactivity condition of pH 5.0. Prostaglandin (PG)1 D synthase (PGDS) is the key enzyme responsible for the formation of PGD 2 and its additional metabolites, the J series of PGs; and these prostanoids exhibit a variety of pharmacological activities and are involved in a number of physiological processes (1). For example, PGD 2 induces vasodilation and bronchoconstriction (2, 3); inhibits platelet aggregation (4); regulates body temperature (5, 6), hormone release (7), and nociception (8); and promotes sleep (9). Furthermore, ⌬ 12 -PGJ 2 inhibits the growth of various tumor cell lines (10), and 15-deoxy-⌬ 12,14 -PGJ 2 is an endogenous ligand for a nuclear receptor, peroxisome proliferator-activated receptor ␥ (11, 12).Two distinct types of PGDS have been isolated and characterized biochemically and immunologically; one is GSHdependent or hematopoietic PGDS (13,14), and the other is GSH-independent or lipocalin-type PGDS (15). Hematopoietic PGDS expressed in antigen-presenting, Kupffer, dendritic, Langerhans (16), megakaryoblastic (17), and mast (18, 19) cells of various organs is involved in the production of PGD 2 as an allergic and inflammatory mediator. In addition, it was characterized as a member of the glutathione S-transferase (GST) gene family (20), whose members are known to catalyze the conjugation of GSH to an electrophilic substrate (21).Recently, we cloned the cDNA for rat hematopoietic PGDS, crystallized the recombinant enzyme, and determined by x-ray crystallography the three-dimensional structure of the enzyme complexed with GSH (22). The deduced amino acid sequence and the tertiary structure of hematopoietic PGDS classified the enzyme as a member of the Sigma class of GSTs. The enzyme isomerizes PGH 2 to PGD 2 sele...

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confidence: 53%

Structural Basis of Hematopoietic Prostaglandin D Synthase Activity Elucidated by Site-directed Mutagenesis

Pinzar¹,

Miyano²,

Kanaoka³

et al. 2000

Journal of Biological Chemistry

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“…Recent crystal structures of the human class pi GST have provided detail of a further binding site located in the hydrophobic cavity between the secondary structural elements β2 and A1. One molecule of N-(2-hydroxethyl)piperazine-N′-2-ethanesulphonic acid (Hepes) and N-morpholino-ethansulphonic acid (Mes) has been shown to bind per GST subunit [29,39]. Competitive binding studies between BSP and ANS, and BSP and Hepes in human GST A1-1 indicate that BSP most probably competes for the Hepes binding site.…”

Section: Discussionmentioning

confidence: 99%

Aflatoxin B1 and sulphobromophthalein binding to the dimeric human glutathione S‐transferase A1‐1 : a fluorescence spectroscopic analysis

Sluis‐Cremer

Wallace

Burke

et al. 1998

European Journal of Biochemistry

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The binding interactions between dimeric human class alpha glutathione S-transferase A1-1 (GST A1-1) and aflatoxin B1 or sulphobromophthalein (BSP) were characterised. Aflatoxin B1 binds to GST A1-1 with a stoichiometry of 1.1 mol/mol of dimeric enzyme. The binding interaction, which can be described by a hyperbolic saturation isotherm (K d ϭ 8Ϯ 2 µM), does not induce major structural changes in the enzyme, nor does it inhibit enzymatic activity. The average distance between the single tryptophan residue (Trp20) of GST A1-1 and protein-bound aflatoxin B1 was calculated to be 22.7 Å by means of fluorescence resonance energy transfer. The aflatoxin-binding region, according to this calculated distance, was determined to be located in the dimer interface cleft near the crystallographic two-fold axis. Hill-plot analyses suggest that a positive co-operative interaction exists between BSP and the dimeric GST A1-1 (h ϭ 1.6 Ϯ0.1 ; K′ ϭ 14Ϯ0.6 µM). The binding of BSP induces a conformational change in the enzyme which is accompanied by a decrease in the molecular flexibility and in the solvent-accessible properties of the enzyme's Trp20 residue. Site-directed mutagenesis of Trp20 (Trp20→Phe) confirms that this residue is situated in the binding environment and although it is not essential for BSP binding, it is involved in the interaction. Furthermore, the structural change associated with BSP binding alters the hyperbolic character of the glutathione saturation curve. This indicates that there may also be a cooperative interaction between glutathione and BSP or that BSP binding induces asymmetric functioning of the two enzyme subunits so that they become unequal in catalytic activity.Keywords : glutathione S-transferase ; aflatoxin B1; sulphobromophthalein; co-operative binding.Glutathione S-transferases (GSTs) are a family of multifunctional enzymes found in all vertebrates, plants, insects, nematodes, yeast and aerobic bacteria. They constitute a complex supergene family that collectively metabolises a broad variety of potentially toxic xenobiotics and reactive endogenous compounds resulting from oxidative metabolism. GSTs are an integral part of the phase-I detoxification mechanism and primarily catalyse the nucleophilic addition of the thiol of reduced glutathione (γ-glutamyl-cysteinyl-glycine) to electrophilic centres in a wide variety of hydrophobic electrophiles, including alkyl and aryl halides, epoxides, quinones and activated alkenes [1]. The dimeric cytosolic GSTs exhibit multiple enzyme forms that, according to primary structure and molecular recognition, can be grouped into one of six species independent gene classes (alpha, kappa, mu, pi, theta and sigma). In addition to their catalytic activities, GSTs also function as ligand binding proteins and thereby facilitate the intracellular storage of a variety of hydrophobic non-substrate compounds, including hormones, metabolites and drugs [2]. The binding of these compounds to GSTs prevents the build up of apolar molecules at lipophilic sites such as membranes...

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“…This was the first structure in which ligand binding outside of the G-and H-sites was demonstrated in a cytosolic GST. Several structures of the human pi-class GST feature binding of the buffersubstances MES or HEPES at a site adjacent to W28, located at the N-terminal end of strand β2 (Ji et al 1997;Oakley et al 1997b;Prade et al 1997) (Figure 8c, d). Later studies demonstrated binding of ligands in various binding sites: Sulfasalazine (a sulfa-drug), cibacron blue (a sulfonated triazine dye) and bromosulfophthalein (used in liver function tests) bind in the H-site (Oakley et al 1999).…”

Section: Gsts As Ligandinsmentioning

confidence: 99%

Glutathione transferases: a structural perspective

Oakley

2011

Drug Metabolism Reviews

317

240

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The glutathione transferases (GSTs) are one of the most important families of detoxifying enzymes in nature. The classic activity of the GSTs is conjugation of compounds with electrophilic centers to the tripeptide glutathione (GSH), but many other activities are now associated with GSTs, including steroid and leukotriene biosynthesis, peroxide degradation, double-bond cis-trans isomerization, dehydroascorbate reduction, Michael addition, and noncatalytic "ligandin" activity (ligand binding and transport). Since the first GST structure was determined in 1991, there has been an explosion in structural data across GSTs of all three families: the cytosolic GSTs, the mitochondrial GSTs, and the membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG family). In this review, the major insights into GST structure and function will be discussed. AbstractThe glutathione transferases (GSTs) are one of the most important families of detoxifying enzymes in nature. The classic activity of the GSTs is conjugation of compounds with electrophilic centers to the tripeptide glutathione (GSH), but many other activities are now associated with GSTs, including steroid and leukotriene biosynthesis, peroxide degradation, double-bond cis-trans isomerization, dehydroascorbate reduction, Michael addition, and non-catalytic "ligandin" activity (ligand binding and transport). Since the first GST structure was determined in 1991, there has been an explosion in structural data across GSTs of all three families: the cytosolic GSTs, the mitochondrial GSTs and the membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG family). In this review, the major insights into GST structure and function will be discussed.

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Structures of class pi glutathione S-transferase from human placenta in complex with substrate, transition-state analogue and inhibitor

Cited by 76 publications

References 35 publications

Structural Basis of Hematopoietic Prostaglandin D Synthase Activity Elucidated by Site-directed Mutagenesis

Structural Basis of Hematopoietic Prostaglandin D Synthase Activity Elucidated by Site-directed Mutagenesis

Aflatoxin B1 and sulphobromophthalein binding to the dimeric human glutathione S‐transferase A1‐1 : a fluorescence spectroscopic analysis

Glutathione transferases: a structural perspective

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