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1.
Glutathione acts as a protein disulphide reductant, which detoxifies herbicides by conjugation, either spontaneously or by the activity of one of a number of glutathione-S-transferases (GSTs), and regulates gene expression in response to environmental stress and pathogen attack. GSTs play roles in both normal cellular metabolisms as well as in the detoxification of a wide variety of xenobiotic compounds, and they have been intensively studied with regard to herbicide detoxification in plants. A newly discovered plant GST subclass has been implicated in numerous stress responses, including those arising from pathogen attack, oxidative stress and heavy-metal toxicity. In addition, plants GSTs play a role in the cellular response to auxins and during the normal metabolism of plant secondary products like anthocyanins and cinnamic acid. The present work involves two in silico analytical approaches—general secondary structure prediction studies of the proteins and detailed signature pattern studies of some selected GST classes in Arabdiopsis thaliana, mustard, maize and bread wheat by standard Bioinformatics tools; structure prediction tools; signature pattern tools; and the evolutionary trends were analyzed by ClustalW. For this purpose, sequences were obtained from standard databases. The work reveals that these proteins are mainly alpha helical in nature with specific signature pattern similar to phosphokinase C, tyrosine kinase and casein kinase II proteins, which are closely related to plant oxidative stress. This study aims to comprehend the relationship of GST gene family and plant oxidative stress with respect to certain specific conserved motifs, which may help in future studies for screening of biomodulators involved in plant stress metabolism.  相似文献   

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Glutathione transferases (GSTs) are ubiquitous, multifunctional proteins encoded by large gene families. In different plant species this gene family is comprised of 25–60 members, that can be grouped into six classes on the basis of sequence identity, gene organization and active site residues in the protein. The Phi and Tau classes are the most represented and are plant specific, while Zeta and Theta GSTs are found also in animals. Despite pronounced sequence and functional diversification, GSTs have maintained a highly conserved three-dimensional structure through evolution. Most GSTs are cytosolic and active as dimers, performing diverse catalytic as well as non-catalytic roles in detoxification of xenobiotics, prevention of oxidative damage and endogenous metabolism. Among their catalytic activities are the conjugation of electrophilic substrates to glutathione, glutathione-dependent isomerizations and reductions of toxic organic hydroperoxides. Their main non-catalytic role is as hormone and flavonoid ligandins. GST genes are predominantly organized in clusters non-randomly distributed in the genome. Phylogenetic studies indicate that plant GSTs have mainly evolved after the divergence of plants, the two prevalent Phi and Tau classes being the result of recent, multiple duplication events.  相似文献   

3.
Hemachand T  Shaha C 《FEBS letters》2003,538(1-3):14-18
On the sperm surface, glutathione S-transferases (GSTs) exist as oocyte binding proteins but their detoxification function in this unique cell type is not known. Using H(2)O(2)- and 4-hydroxynonenal-induced sperm dysfunction models, this study demonstrates that the sperm surface GSTs are able to use extracellular reduced glutathione to inhibit the loss of functional competence of goat spermatozoa; however, in the presence of GST inhibitors, they are unable to do so. In the context of susceptibility of spermatozoa to oxidative stress, this finding that strategically located sperm surface GSTs are important for maintaining the functional competence of sperm is relevant to studies on male infertility.  相似文献   

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Glutathione S-transferases (GSTs) are a family of multi-functional enzymes that play critical roles in the detoxification of xenobiotics and the protection of tissues against oxidative damage. GSTs are important enzymes in plant responses to a number of environmental stresses including herbicides and pathogen attack. Ocs elements are a group of related, 20 bp promoter elements which have been exploited by some plant pathogens to express genes in plants. Ocs elements have also been found to regulate the expression of a plant GST promoter. An Arabidopsis GST gene, called GST6 has been isolated. GST6 expression is under tissue-specific control and is induced following treatment with auxin, salicylic acid and H2O2. The GST6 promoter contains a binding site for two Arabidopsis o cs element b inding f actors (OBF), that has some sequence homology to ocs element sequences. Interestingly, OBP1 (O BF b inding p rotein), a DNA-binding protein that was isolated by screening an Arabidopsis cDNA library with a labeled OBF protein as a probe, binds next to the OBF-binding site on the GST6 promoter. OBP1 was able to significantly stimulate the binding of OBF proteins to the GST6 promoter, raising the possibility that interactions between the OBP1 and OBF proteins may be important for GST6 expression.  相似文献   

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The use of plants to reclaim contaminated soils and groundwater, known as phytoremediation, is a promising biotechnological strategy which has gained a lot of attention in the last few years. Plants have evolved sophisticated detoxification systems against the toxin chemicals: following the uptake, the compounds are activated so that certain functional groups can conjugate hydrophilic molecules, such as thiols. The resulting conjugates are recognized by the tonoplast transporters and sequestered into the vacuoles. The xenobiotic conjugation with glutathione is mediated by enzymes which belong to the superfamily of glutathione S-transferases (GSTs) catalyzing the nucleophylic attack of the sulphur of glutathione on the electrophilic groups of the cytotoxic substrates therefore playing a crucial role in their degradation. This study was designed to identify the putative correlation between structural and functional characteristics of plant GST classes belonging to different plant species. Consequently, the protein sequences of the expressed GSTs have been retrieved from UniGene, classified and then analyzed in order to assess the evolutionary trend and to predict secondary structure. Moreover, the fingerprint analysis was performed with SCAN Prosite in the attempt to correlate meaningful signature profile and biological information. The results evidenced that all the soluble GSTs have a tendency to assume the α-helix secondary structure followed by random coil and β-sheet. The fingerprint analysis revealed that specific signature profiles related mainly to protein phosphorylation are in the GST classes of all considered species thus suggesting that they might be subjected to reversible activation by phosphorylation-mediated regulation. This approach provides the knowledge of the relationship between presence of conserved signature profile and biological function in the view of future selection of GSTs which might be employed in either mutagenesis or genetic engineering studies.  相似文献   

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Molecular Biology Reports - Glutathione S-transferases (GSTs) are multifunctional proteins that help in oxidative stress metabolism and detoxification of xenobiotic compounds. Studies pertaining to...  相似文献   

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Glutathione transferases (GSTs) are a family of enzymes that play important roles in stress tolerance and detoxification in plants. The plant GSTs are divided into four classes (phi, tau, zeta and theta), among which tau is the most numerously represented. To date, studies on GSTs in plants have focused largely on crop species. There is extremely little information on the molecular characteristics of GSTs in gymnosperms. Generalization on GST characteristics unique to gymnosperms and the patterns of GST evolution in plants cannot be made before more members of the gene family in conifers are described. In this study we report three new GSTs from Pinus tabulaeformis, Pinus densata and Pinus yunnanensis. Structural and phylogenetic analyses placed these three GSTs in tau class. The tau GST class is subdivided into three clades and this subdivision seems an ancient event that may have pre-dated the gymnosperm and angiosperm split. Sequence analysis revealed a highly conserved N-terminal domain in contrast to a highly variable C-terminal domain. Mutations even outside the critical glutathione-binding site in N-terminal domain can have pronounced effect on GST catalytic property. Thus, sequence similarity does not parallel functional specificity. The high diversity in C-terminal domain determines a wide range of substrate selectivity and specificity among tau GSTs. Thus the a few conserved residues in C-terminal domain seem essential to maintain the structure of the domain and the protein dimer. More extensive data on GST family organization and a thorough gene-by-gene analysis in conifers are needed to advance our understanding of the true diversity and evolution of GST in structure and function in plants.  相似文献   

11.
Roles for glutathione transferases in plant secondary metabolism   总被引:5,自引:0,他引:5  
Plant glutathione transferases (GSTs) are classified as enzymes of secondary metabolism, but while their roles in catalysing the conjugation and detoxification of herbicides are well known, their endogenous functions are largely obscure. Thus, while the presence of GST-derived S-glutathionylated xenobiotics have been described in many plants, there is little direct evidence for the accumulation of similarly conjugated natural products, despite the presence of a complex and dichotomous metabolic pathway which processes these reaction products. The conservation in glutathione conjugating and processing pathways, the co-regulation of GSTs with inducible plant secondary metabolism and biochemical studies showing the potential of these enzymes to conjugate reactive natural products are all suggestive of important endogenous functions. As a framework for addressing these enigmatic functions we postulate that either: (a) the natural reaction products of GSTs are unstable and undergo reversible S-glutathionylation; (b) the conjugation products of GSTs are very rapidly processed to derived metabolites; (c) GSTs do not catalyse conventional conjugation reactions but instead use glutathione as a cofactor rather than co-substrate; or (d) GSTs are non-catalytic and function as transporter proteins for secondary metabolites and their unstable intermediates. In this review, we describe how enzyme biochemistry and informatics are providing clues as to GST function allowing for the critical evaluation of each of these hypotheses. We also present evidence for the involvement of GSTs in the synthesis of sulfur-containing secondary metabolites such as volatiles and glucosinolates, and the conjugation, transport and storage of reactive oxylipins, phenolics and flavonoids.  相似文献   

12.
Glutathione S-transferases (GSTs) are multifunctional proteins and play a role in detoxification of xenobiotics as well as prevention of oxidative damage. This study exogenously overexpressed PtGSTF4 from Populus trichocarpa and its two orthologs from Populus yatungensis and Populus euphratica in Arabidopsis thaliana, respectively. To elucidate the function of three GSTF4 proteins in stress response, we compared germination and seedling growth in transgenic Arabidopsis with salt and drought treatments. All three Populus GSTF4 genes overexpressed Arabidopsis showed enhanced resistance to salt stress and drought. GSTF4 transgenic plants accumulated less hydrogen peroxide and more chlorophylls and decreased levels of lipid peroxidation under salt stress and drought comparing to the mock control plants. The difference observed by GSH and GSSG measurements indicated GSTF4 proteins may involve in glutathione-dependent peroxide scavenging which lead to reduced oxidative damage. The Arabidopsis transformed with the GSTF4 gene form P. euphratica showed higher germination rate and different performance of affecting GSSG contents comparing with the other two orthologous GST genes under NaCl treatment. These results suggested three Populus GSTF4 orthologs may have functional divergence in stress responding. This study provides insights into molecular mechanisms that underlie salt and drought stress tolerance of Phi GSTs and gives evidence for the functional divergence among orthologs in vivo.  相似文献   

13.
Plant glutathione transferases   总被引:5,自引:0,他引:5  
Dixon DP  Lapthorn A  Edwards R 《Genome biology》2002,3(3):reviews300-reviews300410
The soluble glutathione transferases (GSTs, EC 2.5.1.18) are encoded by a large and diverse gene family in plants, which can be divided on the basis of sequence identity into the phi, tau, theta, zeta and lambda classes. The theta and zeta GSTs have counterparts in animals but the other classes are plant-specific and form the focus of this article. The genome of Arabidopsis thaliana contains 48 GST genes, with the tau and phi classes being the most numerous. The GST proteins have evolved by gene duplication to perform a range of functional roles using the tripeptide glutathione (GSH) as a cosubstrate or coenzyme. GSTs are predominantly expressed in the cytosol, where their GSH-dependent catalytic functions include the conjugation and resulting detoxification of herbicides, the reduction of organic hydroperoxides formed during oxidative stress and the isomerization of maleylacetoacetate to fumarylacetoacetate, a key step in the catabolism of tyrosine. GSTs also have non-catalytic roles, binding flavonoid natural products in the cytosol prior to their deposition in the vacuole. Recent studies have also implicated GSTs as components of ultraviolet-inducible cell signaling pathways and as potential regulators of apoptosis. Although sequence diversification has produced GSTs with multiple functions, the structure of these proteins has been highly conserved. The GSTs thus represent an excellent example of how protein families can diversify to fulfill multiple functions while conserving form and structure.  相似文献   

14.
Glutathione S-transferases (GST) in insects play an important role in the detoxification of many substances including allelochemicals from plants. Induction of GST activity in Myzus persicae in response to secondary metabolites from Brassica plants was determined using different host plant species and confirmed using artificial diet with pure allelochemicals added. The 2,4-dinitro-1-iodobenzene (DNIB) was found to be a useful substrate for identifying particular GSTs in insects. GSTs from M. persicae were purified using different affinity chromatography columns and related kinetic parameters were calculated. GST isoenzymes were characterised using electrophoretic methods. Although SDS-PAGE results indicated similarity among the purified enzymes from each affinity column, biochemical studies indicated significant differences in kinetic parameters. Finally, the GST pattern of M. persicae was discussed in terms of insect adaptation to the presence of plant secondary substances such as the glucosinolates and the isothiocyanates, from Brassicaceae host plants.  相似文献   

15.
Glutathione-S-transferases (GSTs) are a superfamily of enzymes that function to catalyze the nucleophilic attack of glutathione on electrophilic groups of a second substrate. GSTs are present in many organs and have been implicated in the detoxification of endogenous alpha, beta unsaturated aldehydes, including 4-hydroxynonenal (HNE). Exogenous GST protects hippocampal neurons against HNE in culture. To test the hypothesis that overexpression of GST in cells would increase resistance to exogenous or endogenous HNE induced by oxidative stress, stable transfectants of SY5Y neuroblastoma cells with GST were established. Stable GST transfectants demonstrated enzyme activities 13.7 times (Clone 1) and 30 times (Clone 2) higher than cells transfected with vector alone. GST transfectants (both Clones 1 and 2) demonstrated significantly (p <.05) increased resistance to ferrous sulfate/hydrogen peroxide (20.9% for Clone 1; 46.5% for Clone 2), amyloid beta-peptide (12.2% for Clone 1; 27.5.% for Clone 2), and peroxynitrite (24.3% for Clone 1; 43.9% for Clone 2), but not to exogenous application of HNE in culture medium. GST transfectants treated with 1,1,4-tris (acetyloxy)nonane, a nontoxic derivative of HNE that is degraded to HNE intracellularly, demonstrated a statistically significant (p <.05) increase in viability in a dose-dependent manner compared with SY5Y cells transfected with vector alone. These results suggest that overexpression of GST increases resistance to endogenous HNE induced by oxidative stress or released in the degradation of 1,1,4-tris (acetyloxy)nonane, but not to exogenous application of HNE.  相似文献   

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Glutathione transferases (GSTs) form a superfamily of multifunctional proteins with essential roles in cellular detoxification processes and endogenous metabolism. The distribution of fungal-specific class A GSTs was investigated in saprotrophic fungi revealing a recent diversification within this class. Biochemical characterization of eight GSTFuA isoforms from Phanerochaete chrysosporium and Coprinus cinereus demonstrated functional diversity in saprotrophic fungi. The three-dimensional structures of three P. chrysosporium isoforms feature structural differences explaining the functional diversity of these enzymes. Competition experiments between fluorescent probes, and various molecules, showed that these GSTs function as ligandins with various small aromatic compounds, derived from lignin degradation or not, at a L-site overlapping the glutathione binding pocket. By combining genomic data with structural and biochemical determinations, we propose that this class of GST has evolved in response to environmental constraints induced by wood chemistry.  相似文献   

18.
以产黄青霉(Penicillium chrysogenum Thom)cDNA为模板,克隆得到一个新的谷胱甘肽转移酶基因PcgstB,其开放阅读框长651bp,编码216个氨基酸的蛋白质。与已知序列进行BLASTp比较显示,该蛋白具有保守的GST结构域,与烟曲霉GstB的序列一致性最高,达65%。将PcgstB与原核表达载体pTrc99A连接得到表达质粒pTrc-gstB,转化大肠杆菌DH5α,经IPTG诱导后获得以可溶形式表达的重组PcGstB蛋白。以1-chloro-2,4-dinitrobenzene(CDNB)为底物检测,确认该蛋白具有GST活性。  相似文献   

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