Sal003, an effective cell-permeable analog, inhibitis the eIF2α phosphatase.

在大鼠中,Sal003 损伤长时记忆的形成.

在小鼠胚胎成纤维细胞中,Sal003通过增加eIF2a的磷酸化,引起多核糖体的解聚。Sal003通过阻断eIF2a磷酸酯酶的活性,来增加eIF2a的磷酸化；Sal003通过引起的eIF2α的磷酸化,增强由细胞毒素诱导的凋亡信号通路。

Catalytic assay: MEK5 protein isolated from the baculovirus expression system is used to measure kinase activity utilizing PKLight ATP Detection Reagent. The assay is performed using 15 nM GST-MEK5 and 0.75 μM ATP in assay buffer consisting of 25 mM Hepes, pH 7.5, 10 mM MgCl2, 50 mM KCl, 0.2% BSA, 0.01% CHAPS, 100 μM Na3VO4, 0.5 mM DTT and 1% DMSO in the presence of varying concentrations of BIX02189. The kinase reaction mixture is incubated for 90 minutes at room temperature followed by addition of 10 μL of ATP detection reagent for 15 minutes. The relative light unit (RLU) signal is measured and the RLU signals are converted to percent of control (POC) values for the determination of IC50 value.