A tangible method to assess native ferroptosis suppressor activity.

CP: Molecular biology FSP1 GPX4 LC-MS/MS RSL3 affinity purification biochemistry cell death drug discovery enzyme assay lipid peroxidation pull-down assay selenocysteine

Journal

Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676

Informations de publication

Date de publication:
23 Feb 2024
Historique:
received: 14 12 2023
revised: 12 01 2024
accepted: 26 01 2024
medline: 25 2 2024
pubmed: 25 2 2024
entrez: 24 2 2024
Statut: aheadofprint

Résumé

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 GPX4

Identifiants

DOI: 10.1016/j.crmeth.2024.100710 PMID: 38401540
pubmed: 38401540
pii: S2667-2375(24)00025-0
doi: 10.1016/j.crmeth.2024.100710
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

100710

Informations de copyright

Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests M.C. is a co-founder and shareholder of ROSCUE Therapeutics GmbH.

Auteurs

Toshitaka Nakamura (T)

Institute of Metabolism and Cell Death, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany.

Junya Ito (J)

Institute of Metabolism and Cell Death, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany; Laboratory of Food Function Analysis, Tohoku University Graduate School of Agricultural Science, Sendai, Miyagi 980-8572, Japan.

André Santos Dias Mourão (ASD)

Institute of Structural Biology, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany.

Adam Wahida (A)

Institute of Metabolism and Cell Death, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany.

Kiyotaka Nakagawa (K)

Laboratory of Food Function Analysis, Tohoku University Graduate School of Agricultural Science, Sendai, Miyagi 980-8572, Japan.

Eikan Mishima (E)

Institute of Metabolism and Cell Death, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany; Division of Nephrology, Rheumatology and Endocrinology, Tohoku University Graduate School of Medicine, Sendai, Miyagi 980-8574, Japan. Electronic address: eikan@med.tohoku.ac.jp.

Marcus Conrad (M)

Institute of Metabolism and Cell Death, Molecular Targets & Therapeutics Center, Helmholtz Zentrum München, 85764 Neuherberg, Bavaria, Germany. Electronic address: marcus.conrad@helmholtz-munich.de.

Classifications MeSH