2024/09/24 by Esben B. Svenningsen, Fatih Demir, Florian Kromm +4 · 2 voices
Biochemistry, Genetics and Molecular Biology · Medicine · #Cancer-related gene regulation #Peptidase Inhibition and Analysis #Ubiquitin and proteasome pathways
paper · doi:10.26434/chemrxiv-2024-4w549
openalex publication_date 2024/09/24 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/22
The oxidation status of N-terminal cysteines directly dictates protein stability via arginylation and proteasomal degradation. However, only a handful of proteins have been shown to be regulated via this pathway. To date, no methods to detect N-terminal cysteine reactivity and abundance has been reported. Here, we demonstrate, for the first time, that N-terminal cysteine targeting probes can be used in living cells to bind and quantify N-terminal cysteines, discriminating their oxidation state. Using these probes, we identify hundreds of N-terminal cysteines and show that changes in reactivity and abundance under hypoxia can be directly detected. We believe the use of these types of probes will significantly expand our knowledge of this important proteolytic pathway.