Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, Building 37, Room 3128, Bethesda, MD 20892, USA.
J Struct Biol. 2010 May;170(2):216-25. doi: 10.1016/j.jsb.2010.02.013. Epub 2010 Feb 20.
Previously, we identified an arylstibonic acid, NSC13778 that specifically binds to the basic region of the C/EBPalpha B-ZIP domain and disrupts DNA binding. We now examine a panel of 14 additional arylstibonic acid derivatives of NSC13778 for their ability to inhibit the DNA binding of five B-ZIP dimers (c-Fos|JunD, VBP, C/EBPalpha, C/EBPbeta, and CREB). They show various specificities at inhibiting the DNA binding of five B-ZIP domains. NSC13746 inhibits the DNA binding of C/EBPbeta and CREB at 100nM and promiscuously inhibiting the DNA binding of all five proteins in the 1muM range. Dialysis experiments indicate that NSC 13746 binding to the B-ZIP domain is reversible. Thermal denaturation studies indicate that NSC13746 binds the B-ZIP domain. Some compounds specifically inhibit DNA binding, with VBP and c-Fos|JunD being most easily disrupted. These compounds inhibit, with similar specificities to the pure B-ZIP domains, the DNA binding of nuclear extract to the AP1 DNA sequence but no inhibition is observed to SP1 containing oligonucleotide. Transient transfection assays indicate that NSC13746 can inhibit the TPA induced activation of two B-ZIP dependent reporters. These experiments suggest that arylstibonic acids are promising leads for inhibiting the DNA binding of a group of B-ZIP proteins in cells.
先前，我们鉴定了一种芳基膦酸化合物 NSC13778，它可以特异性结合 C/EBPalpha B-ZIP 结构域的碱性区域并破坏 DNA 结合。我们现在检查了 NSC13778 的另外 14 种芳基膦酸衍生物，以研究它们抑制五个 B-ZIP 二聚体（c-Fos|JunD、VBP、C/EBPalpha、C/EBPbeta 和 CREB）的 DNA 结合的能力。它们在抑制五个 B-ZIP 结构域的 DNA 结合方面具有不同的特异性。NSC13746 在 100nM 时抑制 C/EBPbeta 和 CREB 的 DNA 结合，并在 1μM 范围内杂乱无章地抑制所有五种蛋白质的 DNA 结合。透析实验表明 NSC13746 与 B-ZIP 结构域的结合是可逆的。热变性研究表明 NSC13746 结合 B-ZIP 结构域。一些化合物特异性抑制 DNA 结合，其中 VBP 和 c-Fos|JunD 最容易被破坏。这些化合物以与纯 B-ZIP 结构域相似的特异性抑制核提取物与 AP1 DNA 序列的 DNA 结合，但对含有 SP1 的寡核苷酸没有抑制作用。瞬时转染实验表明，NSC13746 可以抑制 TPA 诱导的两个 B-ZIP 依赖性报告基因的激活。这些实验表明，芳基膦酸是抑制细胞中一组 B-ZIP 蛋白的 DNA 结合的有前途的先导化合物。
