5 research outputs found
Structures of relevant compounds.
<p><b>Panel A.</b> Structure of SJ-172550 (<b>1</b>) and a non-reactive analog (<b>2</b>). <b>Panel B.</b> The potential mechanism of covalent adduct formation.</p
Inhibition of MDMX-p53 peptide binding by compound 1 (IC<sub>50</sub> = 3 µM), compound 2 (IC<sub>50</sub>>100 uM).
<p>Inhibition of MDMX-p53 peptide binding by compound 1 (IC<sub>50</sub> = 3 µM), compound 2 (IC<sub>50</sub>>100 uM).</p
Thermal stability equilibria of MDMX.
<p><b>Panel a.</b> Thermal shift data for MDMX (23–111) showing a 7 degree stabilization of the protein’s melting point by addition of compound <b>1</b>. The panel shows individual data sampling points from 3 independent experiments from each condition. <b>Panel b.</b> Dose dependency and time dependency of the effect showing an apparent EC<sub>50</sub> of roughly 1 µM and minimal time dependency. <b>Panel c.</b> Dose dependent reversal of the effects of compound <b>1</b> by TCEP. <b>Panel d.</b> Dose dependent reversal of the effects of compound <b>1</b> by DTT.</p
Formation of covalent adducts between compound 1 and MDMX.
<p><b>Panel a.</b> Mass spectrum arising from unmodified hMDMX (GST-tagged screening construct) showing unmodified mass of the protein. <b>Panel b.</b> Mass spectrum arising from treatment of 20 µM GST-hMDMX with 100 µM of compound <b>1</b> demonstrating multiple alkylation events. Note that 100 µM is well above the solubility limit of compound <b>1</b> and significant aggregation of compound exists. <b>Panel c.</b> Mass spectrum arising from treatment of 1 µM GST-hMDMX with 5 µM of compound <b>1</b> demonstrating no alkylation events. <b>Panel d.</b> Mass spectrum arising from unmodified hMDMX (untagged aa 23 to 111 construct) showing unmodified mass of the protein. <b>Panel e.</b> Mass spectrum arising from treatment of 20 µM hMDMX with 100 µM of compound <b>1</b> demonstrating partial alkylation. <b>Panel f.</b> Mass spectrum arising from treatment of 1 µM hMDMX with 5 µM of compound <b>1</b> demonstrating no alkylation.</p
Reversibility of the interaction of compound 1 with MDMX.
<p><b>Panel a.</b> SPR study of the binding of <b>1</b> (100 µM) to hMDMX (aa 23–111) under non-reducing conditions. While the off-rate is slow, the interaction is reversible. <b>Panel b.</b> SPR study of the binding of <b>1</b> (100 µM) to hMDMX (aa 23–111) under reducing conditions. No binding is observed.</p