Analysis of Protein Degradation in Ferroptosis

The ubiquitin-proteasome system (UPS) is a highly conserved cellular mechanism that degrades and recycles proteins in eukaryotic cells. It involves the tagging of specific target proteins with ubiquitin, a small regulatory protein, which marks them for degradation by the proteasome, a large protein complex that acts as a molecular shredder. Dysfunction of the UPS has been implicated in a wide range of diseases, including cancer, neurodegenerative disorders, and viral infections. Therefore, targeting the UPS has become an attractive therapeutic strategy for many diseases. Ferroptosis is an iron-dependent cell death process that is regulated by multiple levels, including protein degradation. In this chapter, we introduce the detection of UPS-mediated protein degradation in ferroptosis using several techniques such as western blotting, co-immunoprecipitation, in vitro ubiquitination assay, and proteasome assay.