Effects of Substrate-Binding Site Residues on the Biochemical Properties of a Tau Class Glutathione -Transferase from

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2019 Dec 28

PMID 31878175

Citations 4

Authors

Xue Yang

Jinchi Wei

Zhihai Wu

Jie Gao

Affiliations

Soon will be listed here.

Abstract

Glutathione -transferases (GSTs)-an especially plant-specific tau class of GSTs-are key enzymes involved in biotic and abiotic stress responses. To improve the stress resistance of crops via the genetic modification of GSTs, we predicted the amino acids present in the GSH binding site (G-site) and hydrophobic substrate-binding site (H-site) of OsGSTU17, a tau class GST in rice. We then examined the enzyme activity, substrate specificity, enzyme kinetics and thermodynamic stability of the mutant enzymes. Our results showed that the hydrogen bonds between Lys42, Val56, Glu68, and Ser69 of the G-site and glutathione were essential for enzyme activity and thermal stability. The hydrophobic side chains of amino acids of the H-site contributed to enzyme activity toward 4-nitrobenzyl chloride but had an inhibitory effect on enzyme activity toward 1-chloro-2,4-dinitrobenzene and cumene hydroperoxide. Different amino acids of the H-site had different effects on enzyme activity toward a different substrate, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. Moreover, Leu112 and Phe162 were found to inhibit the catalytic efficiency of OsGSTU17 to 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, while Pro16, Leu112, and Trp165 contributed to structural stability. The results of this research enhance the understanding of the relationship between the structure and function of tau class GSTs to improve the abiotic stress resistance of crops.

Citing Articles

Conserved Residues Lys64 and Glu78 at the Subunit Surface of Tau Glutathione Transferase in Rice Affect Structure and Enzymatic Properties.

Yang X, Zhang Z, Wu L, Yang M, Li S, Gao J Int J Mol Sci. 2024; 25(1).

PMID: 38203568 PMC: 10778600. DOI: 10.3390/ijms25010398.

SiteMotif: A graph-based algorithm for deriving structural motifs in Protein Ligand binding sites.

Sankar S, Chandra N PLoS Comput Biol. 2022; 18(2):e1009901.

PMID: 35202398 PMC: 8903255. DOI: 10.1371/journal.pcbi.1009901.

Genome-wide identification and expression profiling of glutathione S-transferase family under multiple abiotic and biotic stresses in Medicago truncatula L.

Hasan M, Singh V, Islam S, Islam M, Ahsan R, Kaundal A PLoS One. 2021; 16(2):e0247170.

PMID: 33606812 PMC: 7894904. DOI: 10.1371/journal.pone.0247170.

Genetic Improvement of Cereals and Grain Legumes.

Nawaz M, Chung G Genes (Basel). 2020; 11(11).

PMID: 33113769 PMC: 7692374. DOI: 10.3390/genes11111255.

References

Edwards R, Dixon D . Plant glutathione transferases. Methods Enzymol. 2006; 401:169-86. DOI: 10.1016/S0076-6879(05)01011-6. View

Xu J, Xing X, Tian Y, Peng R, Xue Y, Zhao W . Transgenic Arabidopsis Plants Expressing Tomato Glutathione S-Transferase Showed Enhanced Resistance to Salt and Drought Stress. PLoS One. 2015; 10(9):e0136960. PMC: 4556630. DOI: 10.1371/journal.pone.0136960. View

Lo Cicero L, Madesis P, Tsaftaris A, Lo Piero A . Tobacco plants over-expressing the sweet orange tau glutathione transferases (CsGSTUs) acquire tolerance to the diphenyl ether herbicide fluorodifen and to salt and drought stresses. Phytochemistry. 2015; 116:69-77. DOI: 10.1016/j.phytochem.2015.03.004. View

Yang Q, Liu Y, Zeng Q . Biochemical functions of the glutathione transferase supergene family of Larix kaempferi. Plant Physiol Biochem. 2014; 77:99-107. DOI: 10.1016/j.plaphy.2014.02.003. View

Liu Y, Han X, Ren L, Yang H, Zeng Q . Functional divergence of the glutathione S-transferase supergene family in Physcomitrella patens reveals complex patterns of large gene family evolution in land plants. Plant Physiol. 2012; 161(2):773-86. PMC: 3561018. DOI: 10.1104/pp.112.205815. View

Barozzi F, Di Sansebastiano G, Sabella E, Aprile A, Piro G, De Bellis L . Glutathione S-transferase related detoxification processes are correlated with receptor-mediated vacuolar sorting mechanisms. Plant Cell Rep. 2017; 36(9):1361-1373. DOI: 10.1007/s00299-017-2159-3. View

Zhu J, Li H, Guo D, Wang Y, Dai H, Mei W . Transcriptome-wide identification and expression analysis of glutathione S-transferase genes involved in flavonoids accumulation in Dracaena cambodiana. Plant Physiol Biochem. 2016; 104:304-11. DOI: 10.1016/j.plaphy.2016.05.012. View

Habig W, Pabst M, Jakoby W . Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974; 249(22):7130-9. View

Ricci G, Caccuri A, Lo Bello M, Pastore A, Piemonte F, Federici G . Colorimetric and fluorometric assays of glutathione transferase based on 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. Anal Biochem. 1994; 218(2):463-5. DOI: 10.1006/abio.1994.1209. View

10.

Dirr H, Reinemer P, Huber R . X-ray crystal structures of cytosolic glutathione S-transferases. Implications for protein architecture, substrate recognition and catalytic function. Eur J Biochem. 1994; 220(3):645-61. DOI: 10.1111/j.1432-1033.1994.tb18666.x. View

11.

Oakley A . Glutathione transferases: a structural perspective. Drug Metab Rev. 2011; 43(2):138-51. DOI: 10.3109/03602532.2011.558093. View

12.

Kapoli P, Axarli I, Platis D, Fragoulaki M, Paine M, Hemingway J . Engineering sensitive glutathione transferase for the detection of xenobiotics. Biosens Bioelectron. 2008; 24(3):498-503. DOI: 10.1016/j.bios.2008.06.037. View

13.

Neuefeind T, Reinemer P, Bieseler B . Plant glutathione S-transferases and herbicide detoxification. Biol Chem. 1997; 378(3-4):199-205. View

14.

Johnson W, Liu S, Ji X, Gilliland G, Armstrong R . Tyrosine 115 participates both in chemical and physical steps of the catalytic mechanism of a glutathione S-transferase. J Biol Chem. 1993; 268(16):11508-11. View

15.

Zeng Q, Wang X . Catalytic properties of glutathione-binding residues in a tau class glutathione transferase (PtGSTU1) from Pinus tabulaeformis. FEBS Lett. 2005; 579(12):2657-62. DOI: 10.1016/j.febslet.2005.03.086. View

16.

Liu H, Tang Z, Han X, Yang Z, Zhang F, Yang H . Divergence in Enzymatic Activities in the Soybean GST Supergene Family Provides New Insight into the Evolutionary Dynamics of Whole-Genome Duplicates. Mol Biol Evol. 2015; 32(11):2844-59. PMC: 4651234. DOI: 10.1093/molbev/msv156. View

17.

Dixon D, Lapthorn A, Edwards R . Plant glutathione transferases. Genome Biol. 2002; 3(3):REVIEWS3004. PMC: 139027. DOI: 10.1186/gb-2002-3-3-reviews3004. View

18.

Edwards R, Dixon D, Walbot V . Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci. 2000; 5(5):193-8. DOI: 10.1016/s1360-1385(00)01601-0. View

19.

Yang X, Sun W, Liu J, Liu Y, Zeng Q . Biochemical and physiological characterization of a tau class glutathione transferase from rice (Oryza sativa). Plant Physiol Biochem. 2009; 47(11-12):1061-8. DOI: 10.1016/j.plaphy.2009.07.003. View

20.

Cummins I, Dixon D, Freitag-Pohl S, Skipsey M, Edwards R . Multiple roles for plant glutathione transferases in xenobiotic detoxification. Drug Metab Rev. 2011; 43(2):266-80. DOI: 10.3109/03602532.2011.552910. View