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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
Author(s) -
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.
Publication year - 2021
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.374
H-Index - 138
ISSN - 2059-7983
DOI - 10.1107/s2059798320016125
Subject(s) - selenocysteine , selenoprotein , covalent bond , chemistry , active site , gpx1 , stereochemistry , gpx4 , biochemistry , glutathione peroxidase , enzyme , glutathione , cysteine , organic chemistry
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.