Comparative Analysis of Two Stress-Inducible tau Class Glutathione Transferases from Glycine max Revealed Significant Catalytic and Structural Diversification

Pouliou, Fotini; Perperopoulou, Fereniki; Labrou, Nikolaos E.

doi:10.2174/0929866524666171026125300

Cited by 5 publications

(6 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The 3-D models of all these eight CmGSTs chiefly contain α helix, β sheet, turn, and random coil ( Figure 6 ). The helices observed in the C-terminal domain contribute to the formation of the H-site and, therefore, to the xenobiotic substrate specificity ( Pouliou et al, 2017 ). Intriguingly, the C-terminal domain was composed mainly of α helices, and the N-terminal domain was composed of α helices and β sheets.…”

Section: Resultsmentioning

confidence: 99%

“…In rice, 79 GST genes were identified, and many of the GST genes were commonly regulated during various abiotic (20), arsenate (32), and biotic stress (48) conditions (Jain et al, 2010). Pouliou et al (2017) find that GmGSTU5-5 and GmGSTU8-8 exhibited high activities as glutathione peroxidases, capable of reducing toxic hydroperoxides in Glycine max. Wang et al (2019) report that 14 of 330 TaGST genes could respond to different abiotic stresses and hormones, especially salt stress and abscisic acid in wheat.…”

Section: Identification Of Hami Melon Gstsmentioning

confidence: 94%

See 1 more Smart Citation

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress

et al. 2021

View full text Add to dashboard Cite

As a group of multifunctional enzymes, glutathione S-transferases (GSTs) participate in oxidative stress resistance and cellular detoxification. Here, we identified 39 CmGST genes with typical binding sites from the Hami melon genome, and they can be classified into seven subfamilies. Their molecular information, chromosomal locations, phylogenetic relationships, synteny relationships, gene structures, protein–protein interactions, structure of 3-D models, and expression levels under cold stress were analyzed. Expression analysis indicates that cold-tolerant Jia Shi-310 (JS) had higher GST enzyme activities and expression levels of 28 stress-related genes under cold stress. Some CmGSTs belonging to Tau, Phi, and DHAR classes play significant roles under cold stress, and they could be regarded as candidate genes for further studies. The present study systematically investigated the characterization of the Hami melon GST gene family, extending our understanding of Hami melon GST mediated stress-response mechanisms in this worldwide fruit.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Identification Of Hami Melon Gstsmentioning

confidence: 94%

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Amino acid sequence alignments and phylogenetic analysis of PvGm GSTUG with the tau class GSTs from G. max and P. vulgaris revealed that the PvGm GSTUG displayed higher identity with the Pv GSTU2-2 and Gm GSTU8-8 isoenzymes and, from the evolutionary point of view, formed a separate clade (Figure 5 ). Although the accurate prediction of the parent sequences was impossible, we can nevertheless speculate that most of the PvGm GSTUG sequence was derived from Pv GSTU2-2 (Chronopoulou et al, 2012a ) and Gm GSTU8-8 (Pouliou et al, 2017 ).…”

Section: Resultsmentioning

confidence: 99%

Expanding the Plant GSTome Through Directed Evolution: DNA Shuffling for the Generation of New Synthetic Enzymes With Engineered Catalytic and Binding Properties

et al. 2018

View full text Add to dashboard Cite

Glutathione transferases (GSTs, EC. 2.5.1.18) are inducible multifunctional enzymes that are essential in the detoxification and degradation of toxic compounds. GSTs have considerable biotechnological potential. In the present work, a new method for the generation of synthetic GSTs was developed. Abiotic stress treatment of Phaseolus vulgaris and Glycine max plants led to the induction of total GST activity and allowed the creation of a GST-enriched cDNA library using degenerated GST-specific primers and reverse transcription-PCR. This library was further diversified by employing directed evolution through DNA shuffling. Activity screening of the evolved library led to the identification of a novel tau class GST enzyme (PvGmGSTUG). The enzyme was purified by affinity chromatography, characterized by kinetic analysis, and its structure was determined by X-ray crystallography. Interestingly, PvGmGSTUG displayed enhanced glutathione hydroperoxidase activity, which was significantly greater than that reported so far for natural tau class GSTs. In addition, the enzyme displayed unusual cooperative kinetics toward 1-chloro-2,4-dinitrochlorobenzene (CDNB) but not toward glutathione. The present work provides an easy approach for the simultaneous shuffling of GST genes from different plants, thus allowing the directed evolution of plants GSTome. This may permit the generation of new synthetic enzymes with interesting properties that are valuable in biotechnology.

show abstract

“…The expression of recombinant CdGSTM1-1 was performed according to the general protocol described in published procedures [19][20][21][22]. E. coli BL21 (DE3) cells harboring recombinant plasmid (pET-3aCdGSTM1-1) were grown at 37 • C in 1 L LB medium containing ampicillin (100 µg/mL).…”

Section: Expression and Purificationmentioning

confidence: 99%

“…The thermal stability of CdGSTM1-1 was also investigated using differential scanning fluorimetry (DSF) on an Applied Biosystems ® real-time PCR StepOne™ instrument, as described by Pouliou et al [19]. Fluorescence monitoring was carried out at 4-99 • C at a rate of 1 • C/min.…”

Section: Thermal Stabilitymentioning

confidence: 99%

Structural and Functional Characterization of Camelus dromedarius Glutathione Transferase M1-1

Perperopoulou

Poudel

Papageorgiou

et al. 2022

Life

Self Cite

View full text Add to dashboard Cite

Glutathione transferases (GSTs; EC. 2.5.1.18) are a large family of multifunctional enzymes that play crucial roles in the metabolism and inactivation of a broad range of xenobiotic compounds. In the present work, we report the kinetic and structural characterization of the isoenzyme GSTM1-1 from Camelus dromedarius (CdGSTM1-1). The CdGSΤM1-1 was expressed in E. coli BL21 (DE3) and was purified by affinity chromatography. Kinetics analysis showed that the enzyme displays a relative narrow substrate specificity and restricted ability to bind xenobiotic compounds. The crystal structures of CdGSΤM1-1 were determined by X-ray crystallography in complex with the substrate (GSH) or the reaction product (S-p-nitrobenzyl-GSH), providing snapshots of the induced-fit catalytic mechanism. The thermodynamic stability of CdGSTM1-1 was investigated using differential scanning fluorimetry (DSF) in the absence and in presence of GSH and S-p-nitrobenzyl-GSH and revealed that the enzyme’s structure is significantly stabilized by its ligands. The results of the present study advance the understanding of camelid GST detoxification mechanisms and their contribution to abiotic stress adaptation in harsh desert conditions.

show abstract

Comparative Analysis of Two Stress-Inducible tau Class Glutathione Transferases from Glycine max Revealed Significant Catalytic and Structural Diversification

Cited by 5 publications

References 0 publications

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress

Expanding the Plant GSTome Through Directed Evolution: DNA Shuffling for the Generation of New Synthetic Enzymes With Engineered Catalytic and Binding Properties

Structural and Functional Characterization of Camelus dromedarius Glutathione Transferase M1-1

Contact Info

Product

Resources

About