Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation

Abstract Microsomal glutathione transferase 1 (MGST1) is a detoxification enzyme belonging to the Membrane Associated Proteins in Eicosanoid and Glutathione Metabolism (MAPEG) superfamily. Here we have used electron crystallography of two-dimensional crystals in order to determine an atomic model of...

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Autores principales:	Qie Kuang, Pasi Purhonen, Johan Ålander, Richard Svensson, Veronika Hoogland, Jens Winerdal, Linda Spahiu, Astrid Ottosson-Wadlund, Caroline Jegerschöld, Ralf Morgenstern, Hans Hebert
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fe2feb0db1a94044957a8d5cb9db2cb5
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spelling	oai:doaj.org-article:fe2feb0db1a94044957a8d5cb9db2cb52021-12-02T12:32:18ZDead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation10.1038/s41598-017-07912-32045-2322https://doaj.org/article/fe2feb0db1a94044957a8d5cb9db2cb52017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07912-3https://doaj.org/toc/2045-2322Abstract Microsomal glutathione transferase 1 (MGST1) is a detoxification enzyme belonging to the Membrane Associated Proteins in Eicosanoid and Glutathione Metabolism (MAPEG) superfamily. Here we have used electron crystallography of two-dimensional crystals in order to determine an atomic model of rat MGST1 in a lipid environment. The model comprises 123 of the 155 amino acid residues, two structured phospholipid molecules, two aliphatic chains and one glutathione (GSH) molecule. The functional unit is a homotrimer centered on the crystallographic three-fold axes of the unit cell. The GSH substrate binds in an extended conformation at the interface between two subunits of the trimer supported by new in vitro mutagenesis data. Mutation of Arginine 130 to alanine resulted in complete loss of activity consistent with a role for Arginine 130 in stabilizing the strongly nucleophilic GSH thiolate required for catalysis. Based on the new model and an electron diffraction data set from crystals soaked with trinitrobenzene, that forms a dead-end Meisenheimer complex with GSH, a difference map was calculated. The map reveals side chain movements opening a cavity that defines the second substrate site.Qie KuangPasi PurhonenJohan ÅlanderRichard SvenssonVeronika HooglandJens WinerdalLinda SpahiuAstrid Ottosson-WadlundCaroline JegerschöldRalf MorgensternHans HebertNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Qie Kuang Pasi Purhonen Johan Ålander Richard Svensson Veronika Hoogland Jens Winerdal Linda Spahiu Astrid Ottosson-Wadlund Caroline Jegerschöld Ralf Morgenstern Hans Hebert Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
description	Abstract Microsomal glutathione transferase 1 (MGST1) is a detoxification enzyme belonging to the Membrane Associated Proteins in Eicosanoid and Glutathione Metabolism (MAPEG) superfamily. Here we have used electron crystallography of two-dimensional crystals in order to determine an atomic model of rat MGST1 in a lipid environment. The model comprises 123 of the 155 amino acid residues, two structured phospholipid molecules, two aliphatic chains and one glutathione (GSH) molecule. The functional unit is a homotrimer centered on the crystallographic three-fold axes of the unit cell. The GSH substrate binds in an extended conformation at the interface between two subunits of the trimer supported by new in vitro mutagenesis data. Mutation of Arginine 130 to alanine resulted in complete loss of activity consistent with a role for Arginine 130 in stabilizing the strongly nucleophilic GSH thiolate required for catalysis. Based on the new model and an electron diffraction data set from crystals soaked with trinitrobenzene, that forms a dead-end Meisenheimer complex with GSH, a difference map was calculated. The map reveals side chain movements opening a cavity that defines the second substrate site.
format	article
author	Qie Kuang Pasi Purhonen Johan Ålander Richard Svensson Veronika Hoogland Jens Winerdal Linda Spahiu Astrid Ottosson-Wadlund Caroline Jegerschöld Ralf Morgenstern Hans Hebert
author_facet	Qie Kuang Pasi Purhonen Johan Ålander Richard Svensson Veronika Hoogland Jens Winerdal Linda Spahiu Astrid Ottosson-Wadlund Caroline Jegerschöld Ralf Morgenstern Hans Hebert
author_sort	Qie Kuang
title	Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
title_short	Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
title_full	Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
title_fullStr	Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
title_full_unstemmed	Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
title_sort	dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/fe2feb0db1a94044957a8d5cb9db2cb5
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Dead-end complex, lipid interactions and catalytic mechanism of microsomal glutathione transferase 1, an electron crystallography and mutagenesis investigation

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