Abstract
Key message
The relative position of domains is critical for enzymatic properties of tau class glutathione S-transferases, and altering the position of linker far away from the active center affects catalytic property.
Abstract
Glutathione S-transferases (GSTs) are a family of phase II detoxification enzymes whose main function is to improve plant resistance to stresses. To understand the structural effects of tau class GSTs on their function, using OsGSTU17 as an example, we predicted the residues involved in the interactions between its domains and linker region. We further detected the structural changes in mutants and the corresponding changes in terms of substrate activity and kinetic parameters. Four pairs of residues, including Ala14 and Trp165, Arg20 and Tyr154, Glu74 and Arg98, Asp77 and Met87, forming hydrogen bonds and salt bridges were found to play important roles in maintaining the relative position between the domains and linker region inside the protein. The hydrogen bond between Trp165 and Ala14 affected the structural stability has been demonstrated in our previous study. The mutant R20A lost almost all catalytic activity. Interestingly, the mutant E74A exhibited a significant decrease in activity towards 7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole, 1-chloro-2, 4-dinitrobenzene and 4-nitrobenzyl chloride, while its activity towards substrate cumene hydroperoxide remained unchanged. Compared with other mutants, the mutant D77A exhibited decreased affinity to its substrates and increased activity towards 1-chloro-2, 4-dinitrobenzene and cumene hydroperoxide, but its thermodynamic stability did not change significantly. The relative position of individual domain was critical for enzymatic properties, and the linker which is far away from the active site could change the enzymatic properties of GSTs via altering the relative position of the individual domain. Our results provide insights into the relationship between structure and function of tau class GSTs.
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Data archiving statement
The sequences of mutants of OsGSTU17 are available in the Dryad (DOI: https://doi.org/10.5061/dryad.zgmsbcc7p).
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Acknowledgements
This work was funded by the National Key R&D Program of Ministry of Agriculture and Rural Affairs of the People’s Republic of China [2018YFD0300207-2]; Department of Science and Technology of Jilin Province [20190301061NY]. We would like to thank the State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, the Chinese Academy of Sciences for enzyme property testing equipment support. And we would like to thank the Technology Center of Protein Research, Tsinghua University and Dr. Xuemin Han of Chinese Academy of forestry for their help in protein circular dichroism spectra. We would like to thank TopEdit (www.topeditsci.com) for English language editing of this manuscript.
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XY and JG contributed to the study conception and design. ZW advised on experimental design and acquired funding. XY performed most of the experiments and data analyses and writing of the article. XY and JG revised the paper during the paper submission and revision. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Yang, X., Wu, Z. & Gao, J. Effects of conserved Arg20, Glu74 and Asp77 on the structure and function of a tau class glutathione S-transferase in rice. Plant Mol Biol 105, 451–462 (2021). https://doi.org/10.1007/s11103-020-01099-4
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DOI: https://doi.org/10.1007/s11103-020-01099-4