NSC624206

NSC624206 (N-(2-((p-Chlorobenzyl)dithio)ethyl)decylamine hydrochloride) is an asymmetric disulfide small molecule that acts as a selective inhibitor of the ubiquitin-activating enzyme E1 (UBA1). It effectively blocks the in vitro ubiquitination-mediated degradation of the cell cycle regulator p27 and induces the accumulation of p27 protein in the HepG2 liver cancer cell model.

Ubiquitin E1:9 μM

Method: Human hepatocellular carcinoma HepG2 cells were treated with NSC624206 (1-10 μM) for 24 h. Western blotting was performed to detect endogenous p27 protein expression levels.
Result: NSC624206 induced the accumulation of p27 protein in HepG2 cells in a dose-dependent manner [1].
Method: Mink lung epithelial Kip16 cells stably expressing GFP-p27 were treated with NSC624206 (1-10 μM) for 24 h. Fluorescence microscopy and Western blotting were used to detect GFP-p27 expression levels.
Result: NSC624206 treatment induced uniform and stable accumulation of GFP-p27 in Kip16 cells, with a significant increase in GFP-p27 levels observed at 1 μM [1].
Method: In vitro E1/E2 thioester formation reactions were assembled containing fluorescein-labeled ubiquitin, E1, E2, and ATP. NSC624206 (1-50 μM) was added, and the formation of E1-ubiquitin and E2-ubiquitin thioester adducts was detected by non-reducing SDS-PAGE.
Result: NSC624206 inhibited the formation of the E1-ubiquitin thioester bond with an IC50 of approximately 9 μM, without significantly affecting E2 activity [1].