Carraz Maëlle, Zwart Wilbert, Phan Trang, Michalides Rob, Brunsveld Luc
Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany; Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, 5600MB Eindhoven, The Netherlands.
Chem Biol. 2009 Jul 31;16(7):702-11. doi: 10.1016/j.chembiol.2009.06.009.
The interaction of estrogen receptor alpha (ERalpha) with the consensus LXXLL motifs of transcriptional coactivators provides an entry for functional ERalpha inhibition. Here, synthetic cell-permeable LXXLL peptide probes are brought forward that allow evaluation of the interaction of specific recognition motifs with ERalpha in the context of the cell. The probes feature a nona-arginine tag that facilitates cellular entry and induces probe localization in nucleoli. The nucleoli localization provides an explicit tool for evaluating the LXXLL motif interaction with ERalpha. The probes compete with coactivators, bind ERalpha, and recruit it into the nucleoli. The physical inhibition of the ERalpha-coactivator interaction by the probes is shown to be correlated with the inhibition of ERalpha-mediated gene transcription. This chemical biology approach allows evaluating the ERalpha-coactivator interaction and inhibitor binding directly in cells.
雌激素受体α（ERα）与转录共激活因子的共有LXXLL基序之间的相互作用为功能性ERα抑制提供了切入点。在此，提出了可穿透细胞的合成LXXLL肽探针，其能够在细胞环境中评估特定识别基序与ERα的相互作用。这些探针具有一个九聚精氨酸标签，有助于细胞摄取并诱导探针定位于核仁。核仁定位为评估LXXLL基序与ERα的相互作用提供了一个明确的工具。这些探针与共激活因子竞争，结合ERα，并将其招募到核仁中。结果表明，探针对ERα-共激活因子相互作用的物理抑制与ERα介导的基因转录抑制相关。这种化学生物学方法能够直接在细胞中评估ERα-共激活因子的相互作用以及抑制剂的结合情况。
