Three-dimensional structure of glutathione S-transferase from Arabidopsis thaliana at 2.2 Å resolution: Structural characterization of herbicide-conjugating plant glutathione S-transferases and a novel active site architecture

Reinemer, Peter; Prade, Lars; Hof, Peter; Neuefeind, Torsten; Huber, Robert; Zettl, Rolf; Palme, Klaus; Schell, Jeff; Koelln, I.; Bartunik, H.D.; Bieseler, Barbara

doi:10.1006/jmbi.1996.0024

Cited by 201 publications

(240 citation statements)

References 73 publications

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“…The other conserved Arg and Ser are present in these sequences. In the available crystal structure data for an Arabidopsis GST, maize GSTI and maize GSTIII, Glu-67, Ser-68, and Arg-69 (GSTI numbering) are all involved in binding charged groups on the ␥-glutamyl portion of the substrate GSH, as part of the "G-site" (Reinemer et al, 1996;Neuefeind et al, 1997aNeuefeind et al, , 1997b. The complete conservation of Ser-68 in all maize and soybean GSTs is consistent with a critical role in substrate binding.…”

Section: Structural Features Of Gstsmentioning

confidence: 75%

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A Genomics Approach to the Comprehensive Analysis of the Glutathione S-Transferase Gene Family in Soybean and Maize

McGonigle

Keeler²,

Lau³

et al. 2000

Plant Physiology

283

264

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By BLAST searching a large expressed sequence tag database for glutathione S-transferase (GST) sequences we have identified 25 soybean (Glycine max) and 42 maize (Zea mays) clones and obtained accurate full-length GST sequences. These clones probably represent the majority of members of the GST multigene family in these species. Plant GSTs are divided according to sequence similarity into three categories: types I, II, and III. Among these GSTs only the active site serine, as well as another serine and arginine in or near the “G-site” are conserved throughout. Type III GSTs have four conserved sequence patches mapping to distinct structural features. Expression analysis reveals the distribution of GSTs in different tissues and treatments: Maize GSTI is overall the most highly expressed in maize, whereas the previously unknown GmGST 8 is most abundant in soybean. Using DNA microarray analysis we observed increased expression among the type III GSTs after inducer treatment of maize shoots, with different genes responding to different treatments. Protein activity for a subset of GSTs varied widely with seven substrates, and any GST exhibiting greater than marginal activity with chloro-2,4 dinitrobenzene activity also exhibited significant activity with all other substrates, suggesting broad individual enzyme substrate specificity.

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Section: Structural Features Of Gstsmentioning

confidence: 75%

“…Mammalian GSTs are categorized into six classes (alpha, mu, pi, sigma, theta, and zeta) based on amino acid identity, immunocrossreactivity, and substrate specificity. Several plant GSTs have been crystallized and their structure characterized (Reinemer et al, 1996;Neuefeind et al, 1997aNeuefeind et al, , 1997b.…”

mentioning

confidence: 99%

A Genomics Approach to the Comprehensive Analysis of the Glutathione S-Transferase Gene Family in Soybean and Maize

McGonigle

Keeler²,

Lau³

et al. 2000

Plant Physiology

283

264

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“…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). This enzyme, a type I GST from Arabidopsis (pm24; Fig.…”

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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“…Several X-ray structures for GST enzymes of the a, p and ~t classes have become available over the last five years (for reviews, see Dirr et al, 1994 andParker, 1994), followed by structures of GST enzymes from squid (Ji et al,l995), Schistosoma japonicum (Lim et al, 1994;McTigue et al, 1995), by the first X-ray structure of a 8-class GST from the fly Lucilia cuprina , and, most recently, by that of a herbicide-conjugating plant GST from Arubidopsis thaliana (Reinemer et al, 1996), which from its protein sequence falls in the proposed 4 class of GST enzymes (Blocki et al, 1993). As a consequence, the amassed data on substrate specificity and catalytic activity of glutathione S-transferases and mutants thereof, obtained by protein engineering, can now often be discussed in great structural detail.…”

Section: Discussionmentioning

confidence: 99%

Protein Engineering Studies of Dichloromethane Dehalogenase/Glutathione S‐Transferase from Methylophilus Sp. Strain DM11

Vuilleumier

Leistnger

1996

European Journal of Biochemistry

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The structural gene for dichloromethane dehalogenase/glutathione S-transferase (GST, EC 2.5.1.18) from Methylophilus sp. strain DM1 1 was subcloned into a multicopy plasmid under the control of the T7 polymerase promoter, allowing expression in Escherichia coli and easy purification of the enzyme in good yield. Several point mutations leading to amino acid changes at residues Tyr6, His8 and Serl2 of the protein were introduced in this gene. Mutations at Tyr6, the N-terminal tyrosine known to be essential for enzymatic activity in glutathione S-transferases of the a, and n classes, had little effect on the activity of dichloromethane dehalogenase. The same applied for mutations at residue H i d , which from multiple alignments of GST sequences may also correspond to the conserved N-terminal tyrosine residue of GST enzymes. The higher turnover rate of the wild-type enzyme with dibromomethane compared with dichloromethane was lost in mutants with amino acid replacements at residue Hiss, but retained in mutant proteins at Tyr6. Mutations at Serl2 led to mutants with drastically reduced enzymatic activity, pinpointing this residue as an essential determinant of catalytic efficiency.

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Three-dimensional structure of glutathione S-transferase from Arabidopsis thaliana at 2.2 Å resolution: Structural characterization of herbicide-conjugating plant glutathione S-transferases and a novel active site architecture

Cited by 201 publications

References 73 publications

A Genomics Approach to the Comprehensive Analysis of the Glutathione S-Transferase Gene Family in Soybean and Maize

A Genomics Approach to the Comprehensive Analysis of the Glutathione S-Transferase Gene Family in Soybean and Maize

Isolation and Characterization of GlutathioneS-Transferase Isozymes from Sorghum1

Protein Engineering Studies of Dichloromethane Dehalogenase/Glutathione S‐Transferase from Methylophilus Sp. Strain DM11

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