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Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162

Wild-type human glutathione peroxidase 4 (GPX4) was co-expressed with SBP2 (selenocysteine insertion sequence-binding protein 2) in human HEK cells to achieve efficient production of this selenocysteine-containing enzyme on a preparative scale for structural biology. The protein was purified and cry...

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Autores principales: Moosmayer, Dieter, Hilpmann, André, Hoffmann, Jutta, Schnirch, Lennart, Zimmermann, Katja, Badock, Volker, Furst, Laura, Eaton, John K., Viswanathan, Vasanthi S., Schreiber, Stuart L., Gradl, Stefan, Hillig, Roman C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7869902/
https://www.ncbi.nlm.nih.gov/pubmed/33559612
http://dx.doi.org/10.1107/S2059798320016125
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author Moosmayer, Dieter
Hilpmann, André
Hoffmann, Jutta
Schnirch, Lennart
Zimmermann, Katja
Badock, Volker
Furst, Laura
Eaton, John K.
Viswanathan, Vasanthi S.
Schreiber, Stuart L.
Gradl, Stefan
Hillig, Roman C.
author_facet Moosmayer, Dieter
Hilpmann, André
Hoffmann, Jutta
Schnirch, Lennart
Zimmermann, Katja
Badock, Volker
Furst, Laura
Eaton, John K.
Viswanathan, Vasanthi S.
Schreiber, Stuart L.
Gradl, Stefan
Hillig, Roman C.
author_sort Moosmayer, Dieter
collection PubMed
description Wild-type human glutathione peroxidase 4 (GPX4) was co-expressed with SBP2 (selenocysteine insertion sequence-binding protein 2) in human HEK cells to achieve efficient production of this selenocysteine-containing enzyme on a preparative scale for structural biology. The protein was purified and crystallized, and the crystal structure of the wild-type form of GPX4 was determined at 1.0 Å resolution. The overall fold and the active site are conserved compared with previously determined crystal structures of mutated forms of GPX4. A mass-spectrometry-based approach was developed to monitor the reaction of the active-site selenocysteine Sec46 with covalent inhibitors. This, together with the introduction of a surface mutant (Cys66Ser), enabled the crystal structure determination of GPX4 in complex with the covalent inhibitor ML162 [(S)-enantiomer]. The mass-spectrometry-based approach described here opens the path to further co-complex crystal structures of this potential cancer drug target in complex with covalent inhibitors.
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spelling pubmed-78699022021-02-19 Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162 Moosmayer, Dieter Hilpmann, André Hoffmann, Jutta Schnirch, Lennart Zimmermann, Katja Badock, Volker Furst, Laura Eaton, John K. Viswanathan, Vasanthi S. Schreiber, Stuart L. Gradl, Stefan Hillig, Roman C. Acta Crystallogr D Struct Biol Research Papers Wild-type human glutathione peroxidase 4 (GPX4) was co-expressed with SBP2 (selenocysteine insertion sequence-binding protein 2) in human HEK cells to achieve efficient production of this selenocysteine-containing enzyme on a preparative scale for structural biology. The protein was purified and crystallized, and the crystal structure of the wild-type form of GPX4 was determined at 1.0 Å resolution. The overall fold and the active site are conserved compared with previously determined crystal structures of mutated forms of GPX4. A mass-spectrometry-based approach was developed to monitor the reaction of the active-site selenocysteine Sec46 with covalent inhibitors. This, together with the introduction of a surface mutant (Cys66Ser), enabled the crystal structure determination of GPX4 in complex with the covalent inhibitor ML162 [(S)-enantiomer]. The mass-spectrometry-based approach described here opens the path to further co-complex crystal structures of this potential cancer drug target in complex with covalent inhibitors. International Union of Crystallography 2021-01-26 /pmc/articles/PMC7869902/ /pubmed/33559612 http://dx.doi.org/10.1107/S2059798320016125 Text en © Moosmayer et al. 2021 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/
spellingShingle Research Papers
Moosmayer, Dieter
Hilpmann, André
Hoffmann, Jutta
Schnirch, Lennart
Zimmermann, Katja
Badock, Volker
Furst, Laura
Eaton, John K.
Viswanathan, Vasanthi S.
Schreiber, Stuart L.
Gradl, Stefan
Hillig, Roman C.
Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title_full Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title_fullStr Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title_full_unstemmed Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title_short Crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ML162
title_sort crystal structures of the selenoprotein glutathione peroxidase 4 in its apo form and in complex with the covalently bound inhibitor ml162
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7869902/
https://www.ncbi.nlm.nih.gov/pubmed/33559612
http://dx.doi.org/10.1107/S2059798320016125
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