Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>

Arthropod Glutathione S-transferases (GSTs) constitute a large family of multifunctional enzymes that are mainly associated with xenobiotic or stress adaptation. GST-mediated xenobiotic adaptation takes place through direct metabolism or sequestration of xenobiotics, and/or indirectly by providing p...

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Autores principales:	Yanjun Liu, Timothy Moural, Sonu Koirala B K, Jonathan Hernandez, Zhongjian Shen, Andrei Alyokhin, Fang Zhu
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	glutathione S-transferase xenobiotic adaptation enzyme kinetics crystal and co-crystal structures pesticide inhibition conjugation Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/c1231fe8761741e2a3c0c8514bf98e37
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spelling	oai:doaj.org-article:c1231fe8761741e2a3c0c8514bf98e372021-11-11T17:20:16ZStructural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>10.3390/ijms2221119211422-00671661-6596https://doaj.org/article/c1231fe8761741e2a3c0c8514bf98e372021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11921https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Arthropod Glutathione S-transferases (GSTs) constitute a large family of multifunctional enzymes that are mainly associated with xenobiotic or stress adaptation. GST-mediated xenobiotic adaptation takes place through direct metabolism or sequestration of xenobiotics, and/or indirectly by providing protection against oxidative stress induced by xenobiotic exposure. To date, the roles of GSTs in xenobiotic adaptation in the Colorado potato beetle (CPB), a notorious agricultural pest of plants within Solanaceae, have not been well studied. Here, we functionally expressed and characterized an unclassified-class GST, LdGSTu1. The three-dimensional structure of the LdGSTu1 was solved with a resolution up to 1.8 Å by X-ray crystallography. The signature motif VSDGPPSL was identified in the “G-site”, and it contains the catalytically active residue Ser14. Recombinant LdGSTu1 was used to determine enzyme activity and kinetic parameters using 1-chloro-2, 4-dinitrobenzene (CDNB), GSH, p-nitrophenyl acetate (PNA) as substrates. The enzyme kinetic parameters and enzyme-substrate interaction studies demonstrated that LdGSTu1 could catalyze the conjugation of GSH to both CDNB and PNA, with a higher turnover number for CDNB than PNA. The LdGSTu1 enzyme inhibition assays demonstrated that the enzymatic conjugation of GSH to CDNB was inhibited by multiple pesticides, suggesting a potential function of LdGSTu1 in xenobiotic adaptation.Yanjun LiuTimothy MouralSonu Koirala B KJonathan HernandezZhongjian ShenAndrei AlyokhinFang ZhuMDPI AGarticleglutathione S-transferasexenobiotic adaptationenzyme kineticscrystal and co-crystal structurespesticide inhibitionconjugationBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11921, p 11921 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	glutathione S-transferase xenobiotic adaptation enzyme kinetics crystal and co-crystal structures pesticide inhibition conjugation Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	glutathione S-transferase xenobiotic adaptation enzyme kinetics crystal and co-crystal structures pesticide inhibition conjugation Biology (General) QH301-705.5 Chemistry QD1-999 Yanjun Liu Timothy Moural Sonu Koirala B K Jonathan Hernandez Zhongjian Shen Andrei Alyokhin Fang Zhu Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
description	Arthropod Glutathione S-transferases (GSTs) constitute a large family of multifunctional enzymes that are mainly associated with xenobiotic or stress adaptation. GST-mediated xenobiotic adaptation takes place through direct metabolism or sequestration of xenobiotics, and/or indirectly by providing protection against oxidative stress induced by xenobiotic exposure. To date, the roles of GSTs in xenobiotic adaptation in the Colorado potato beetle (CPB), a notorious agricultural pest of plants within Solanaceae, have not been well studied. Here, we functionally expressed and characterized an unclassified-class GST, LdGSTu1. The three-dimensional structure of the LdGSTu1 was solved with a resolution up to 1.8 Å by X-ray crystallography. The signature motif VSDGPPSL was identified in the “G-site”, and it contains the catalytically active residue Ser14. Recombinant LdGSTu1 was used to determine enzyme activity and kinetic parameters using 1-chloro-2, 4-dinitrobenzene (CDNB), GSH, p-nitrophenyl acetate (PNA) as substrates. The enzyme kinetic parameters and enzyme-substrate interaction studies demonstrated that LdGSTu1 could catalyze the conjugation of GSH to both CDNB and PNA, with a higher turnover number for CDNB than PNA. The LdGSTu1 enzyme inhibition assays demonstrated that the enzymatic conjugation of GSH to CDNB was inhibited by multiple pesticides, suggesting a potential function of LdGSTu1 in xenobiotic adaptation.
format	article
author	Yanjun Liu Timothy Moural Sonu Koirala B K Jonathan Hernandez Zhongjian Shen Andrei Alyokhin Fang Zhu
author_facet	Yanjun Liu Timothy Moural Sonu Koirala B K Jonathan Hernandez Zhongjian Shen Andrei Alyokhin Fang Zhu
author_sort	Yanjun Liu
title	Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
title_short	Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
title_full	Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
title_fullStr	Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
title_full_unstemmed	Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>
title_sort	structural and functional characterization of one unclassified glutathione s-transferase in xenobiotic adaptation of <i>leptinotarsa decemlineata</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/c1231fe8761741e2a3c0c8514bf98e37
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Structural and Functional Characterization of One Unclassified Glutathione S-Transferase in Xenobiotic Adaptation of <i>Leptinotarsa decemlineata</i>

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