Crystal structure of a theta-class glutathione transferase.

Wilce, Matthew Charles James; Board, Philip G.; Feil, Susanne; Parker, M.W.

doi:10.1002/j.1460-2075.1995.tb07207.x

Cited by 225 publications

(235 citation statements)

References 48 publications

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“…In addition, agGST1-2 possesses a serine residue near the N terminus (Ser-10) and an asparagine (Asn-48) (shown in bold in Fig. 5) near the invariant proline, both of which are characteristics of insect class I GSTs (22).…”

Section: Resultsmentioning

confidence: 99%

Cloning and Localization of a Glutathione S-transferase Class I Gene from Anopheles gambiae

Ranson

Cornel

Fournier

et al. 1997

Journal of Biological Chemistry

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1,1,1-Trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) resistance in both adults and larvae of Anopheles gambiae is mediated by stage-specific glutathione S-transferases (GSTs). On the basis of their biochemical characteristics the larval resistance-associated GSTs are likely to be insect class I GSTs. Aggst1-2, a class I GST gene, which is expressed in larvae, has been cloned from the malaria vector A. gambiae. The gene was inserted into a bacterial expression system, and the detection of 1-chloro-2,4-dinitrobenzene (CDNB) conjugating activity in Eschericia coli expressing the recombinant enzyme confirmed that aggst1-2 encodes a catalytically active GST. The gene encodes a 209 amino acid protein with 46% sequence similarity to a Drosophila melanogaster class I GST (GST-D1), 44% similarity with a Musca domestica class I GST (MdGST-1), but only low levels of homology with class II insect GSTs, including the adult specific AgGST2-1 from A. gambiae. Southern analysis of genomic DNA indicated that A. gambiae has multiple class I GSTs. In situ hybridization of class I genomic and cDNA clones to polytene chromosomes identified a single region of complementarity on chromosome 2R division 18B, suggesting that these class I GSTs in A. gambiae are arranged sequentially in the genome. Three positive overlapping recombinant clones were identified from an A. gambiae genomic library. Mapping and partial sequencing of these clones suggests that there are several GSTs and truncated GST pseudogenes within the 30kb of DNA that these clones span.

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Section: Resultsmentioning

confidence: 99%

Cloning and Localization of a Glutathione S-transferase Class I Gene from Anopheles gambiae

Ranson

Cornel

Fournier

et al. 1997

Journal of Biological Chemistry

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“…Site-directed mutagenesis studies of the -GST from the Australian sheep blowfly (Lucilia cuprina) and the -GST from the bacterium Methylophilus sp. strain DM11 indicated that the Ser-9 and Ser-12 residues, respectively, are essential for enzyme activity (Board et al, 1995;Vuilleumier and Leisinger, 1996). Furthermore, the three-dimensional structure of a -GST from L. cuprina showed that Ser-9 occupies a position close to that of the catalytically important Tyr of mammalian ␣-, -, and -GSTs (Wilce et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

“…strain DM11 indicated that the Ser-9 and Ser-12 residues, respectively, are essential for enzyme activity (Board et al, 1995;Vuilleumier and Leisinger, 1996). Furthermore, the three-dimensional structure of a -GST from L. cuprina showed that Ser-9 occupies a position close to that of the catalytically important Tyr of mammalian ␣-, -, and -GSTs (Wilce et al, 1995). Recently, the first three-dimensional structure of a plant GST was reported (Reinemer et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of GlutathioneS-Transferase Isozymes from Sorghum1

1998

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Two glutathione S-transferase (GST) isozymes, A1/A1 and B1/B2, were purified from etiolated, O-1,3-dioxolan-2-yl-methyl-2,2,2,-trifluoro-4 -chloroacetophenone-oxime-treated sorghum (Sorghum bicolor L. Moench) shoots. GST A1/A1, a constitutively expressed homodimer, had a subunit molecular mass of 26 kD and an isoelectric point of 4.9. GST A1/A1 exhibited high activity with 1-chloro-2, 4,dinitrobenzene (CDNB) but low activity with the chloroacetanilide herbicide metolachlor. For GST A1/A1, the random, rapidequilibrium bireactant kinetic model provided a good description of the kinetic data for the substrates CDNB and glutathione (GSH). GST B1/B2 was a heterodimer with subunit molecular masses of 26 kD (designated the B1 subunit) and 28 kD (designated the B2 subunit) and a native isoelectric point of 4.8. GST B1/B2 exhibited low activity with CDNB and high activity with metolachlor as the substrate. The kinetics of GST B1/B2 activity with GSH and metolachlor fit a model describing a multisite enzyme having two binding sites with different affinities for these substrates. Both GST A1/A1 and GST B1/B2 exhibited GSH-conjugating activity with ethacrynic acid and GSH peroxidase activity with cumene hydroperoxide, 9-hydroperoxy-trans-10,cis-12-octadecadienoic acid and 13-hydroperoxy-cis-9,trans-11-octadecadienoic acid. Both GST A1/A1 and GST B1/B2 are glycoproteins, as indicated by their binding of concanavalin A. Polyclonal antibodies raised against GST A1/A1 exhibited cross-reactivity with the B1 subunit of GST B1/B2. Comparisons of the N-terminal amino acid sequences of the GST A1, B1, and B2 subunits with other type I -GSTs indicated a high degree of homology with the maize GST I subunit and a sugarcane GST.GSTs (EC 2.5.1.18) are dimeric enzymes found in mammals, insects, plants, and microbes that catalyze nucleophilic attack by the thiolate anion of GSH at electrophilic centers of hydrophobic molecules (Mannervik and Danielson, 1988). In addition to catalyzing GSH conjugation, GSTs also exhibit GSH peroxidase activity and ligandbinding functions (Mannervik and Danielson, 1988;Marrs, 1996). Mammalian GSTs compose a multigene family; in rat liver at least 13 different cytosolic GST subunits are found as either heterodimers or homodimers . Mammalian cytosolic GSTs have been divided into four classes (␣, , , and ) based on immunological, biochemical, and sequence similarities (Buetler and Eaton, 1992). It is well established that mammalian GSTs play an important role in the detoxification of electrophilic xenobiotics (Mannervik and Danielson, 1988). Although endogenous substrates for mammalian GSTs have not been clearly defined, there is evidence that ␣-GSTs protect against oxidative stress by detoxifying reactive products generated by lipid peroxidation (Ålin et al., 1985;Ketterer and Coles, 1991;Singhal et al., 1992).In general, plant GSTs have not been as well characterized as mammalian GSTs. Plant cytosolic GSTs belong to the archaic class of GSTs (Meyer et al., 1991;Marrs, 1996). This class, which is very heterog...

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“…The structure of a Theta-class GST from Lucilia cuprina revealed that the insect GST has a deeper active site than mammalian GSTs. Furthermore, amino acid sequence alignment has suggested that the extended α-helix 5 in the mammalian Theta-class isoenzymes would result in an even deeper active site in these enzymes [26]. Because of the apparent increased affinity of GSTT2-2 for substrates with increased carbon chain length, and the inability of GSTT2-2 to bind to conventional glutathione\agarose affinity matrices, we attempted to purify GSTT2-2 using a GSH-agarose matrix with a very long spacer arm, equivalent to 20 carbon atoms.…”

Section: Figure 5 Ph Profile Of Gstt2-2 With Cumene Hydroperoxide As mentioning

confidence: 99%

Purification and characterization of a recombinant human Theta-class glutathione transferase (GSTT2-2)

Tan

Board

1996

Biochemical Journal

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A cDNA encoding the human Theta-class glutathione transferase GSTT2-2 was expressed in Escherichia coli as a ubiquitin fusion protein. The co-translational removal of the ubiquitin by a cloned ubiquitin-specific protease, Ubp1, generates enzymically active GSTT2-2 without any additional N-terminal residues. The recombinant isoenzyme was purified to apparent homogeneity by DEAE anion-exchange, gel filtration, dye ligand chromatography and high resolution anion-exchange chromatography on Mono Q FPLC. The recombinant enzyme had significant activity with a range of substrates, including cumene hydroperoxide and 1-menapthyl sulphate. The activity of GSTT2-2 with a range of secondary lipid peroxidation products such as the trans,trans-alka-2,4-dienals and trans-alk-2-enals, as well as its glutathione peroxidase activity with organic hydroperoxides, suggest that it may play a significant role in protection against the products of lipid peroxidation.

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Crystal structure of a theta-class glutathione transferase.

Cited by 225 publications

References 48 publications

Cloning and Localization of a Glutathione S-transferase Class I Gene from Anopheles gambiae

Cloning and Localization of a Glutathione S-transferase Class I Gene from Anopheles gambiae

Isolation and Characterization of GlutathioneS-Transferase Isozymes from Sorghum1

Purification and characterization of a recombinant human Theta-class glutathione transferase (GSTT2-2)

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