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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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International Union of Crystallography
2021
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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. |
format | Online Article Text |
id | pubmed-7869902 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
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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