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Nakamura, T. ; Ito, J. ; Mourao, A. ; Wahida, A. ; Nakagawa, K.* ; Mishima, E. ; Conrad, M.

A tangible method to assess native ferroptosis suppressor activity.

Cell Rep. Methods 4, 18:100710 (2024)
DOI PMC
Creative Commons Lizenzvertrag
Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.
Ferroptosis, a regulated cell death hallmarked by unrestrained lipid peroxidation, plays a pivotal role in the pathophysiology of various diseases, making it a promising therapeutic target. Glutathione peroxidase 4 (GPX4) prevents ferroptosis by reducing (phospho)lipid hydroperoxides, yet evaluation of its actual activity has remained arduous. Here, we present a tangible method using affinity-purified GPX4 to capture a snapshot of its native activity. Next to measuring GPX4 activity, this improved method allows for the investigation of mutational GPX4 activity, exemplified by the GPX4U46C mutant lacking selenocysteine at its active site, as well as the evaluation of GPX4 inhibitors, such as RSL3, as a showcase. Furthermore, we apply this method to the second ferroptosis guardian, ferroptosis suppressor protein 1, to validate the newly identified ferroptosis inhibitor WIN62577. Together, these methods open up opportunities for evaluating alternative ferroptosis suppression mechanisms.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Cp: Molecular Biology ; Fsp1 ; Gpx4 ; Lc-ms/ms ; Rsl3 ; Affinity Purification ; Biochemistry ; Cell Death ; Drug Discovery ; Enzyme Assay ; Lipid Peroxidation ; Pull-down Assay ; Selenocysteine; Cancer-cells; Separation; Death; Gpx4
ISSN (print) / ISBN 2667-2375
e-ISSN 2667-2375
Journal Cell Reports Methods
Quellenangaben Volume: 4, Issue: 3, Pages: 18, Article Number: 100710 Supplement: ,
Publisher Elsevier
Publishing Place 50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Metabolism and Cell Death (MCD)
Institute of Structural Biology (STB)
Grants JSPS KAKENHI
CRC TRR 353
German Federal Ministry of Education and Research (BMBF) FERROPATH
European Research Council (ERC)
Deutsche Forschungsgemeinschaft (DFG)