Proteomic Shift in Mouse Embryonic Fibroblasts Pfa1 during Erastin, ML210, and BSO-Induced Ferroptosis

Ferroptosis is a unique variety of non-apoptotic cell death, driven by massive lipid oxidation in an iron-dependent manner. Since ferroptosis was introduced as a concept in 2012, it has demonstrated its essential role in the pathogenesis in neurodegenerative diseases and an important role in therapy-resistant cancer cells. Thus, detailed molecular understanding of both canonical and alternative ferroptosis pathways is required. There is a set of widely used chemical agents to modulate ferroptosis using different pathway targets: erastin blocks cystine-glutamate antiporter, system xc(-); ML210 directly inactivates GPX4; and L-buthionine sulfoximine (BSO) inhibits & gamma;-glutamylcysteine synthetase, an essential enzyme for glutathione synthesis de novo. Most studies have focused on the lipidomic profiling of model systems undergoing death in a ferroptotic modality. In this study, we developed high-quality shotgun proteome sequencing during ferroptosis induction by three widely used chemical agents (erastin, ML210, and BSO) before and after 24 and 48 h of treatment. Chromato-mass spectra were registered in DDA mode and are suitable for further label-free quantification. Both processed and raw files are publicly available and could be a valuable dynamic proteome map for further ferroptosis investigation. Dataset: Shotgun proteomics are available at the PRoteomics IDEntifications (PRIDE) Archive database with the following dataset identifiers: Erastin experiment-PXD041463 (https://www.ebi.ac.uk/pride/archive/projects/PXD041463, DOI: 10.6019/PXD041463); ML210 experiment-PXD041327 (https://www.ebi.ac.uk/pride/archive/projects/PXD041327, DOI: 10.6019/PXD041327); BSO-PXD041415 (https://www.ebi.ac.uk/pride/archive/projects/PXD041415, DOI: 10.6019/PXD041415). Dataset License: CC-BY-NC

Automated summary

Ferroptosis is a form of non-apoptotic cell death that involves lipid oxidation driven by iron. It plays critical roles in the pathogenesis of neurodegenerative diseases and therapy-resistant cancer cells. This study developed a high-quality shotgun proteome sequencing method to analyze the effects of three chemical agents on ferroptosis induction.