Müller Judith, Schust Jochen, Berg Thorsten
Max Planck Institute of Biochemistry, Department of Molecular Biology, and Munich Center for Integrated Protein Science (CIPSM), Am Klopferspitz 18, 82152 Martinsried, Germany.
Anal Biochem. 2008 Apr 15;375(2):249-54. doi: 10.1016/j.ab.2008.01.017. Epub 2008 Jan 18.
Signal transducer and activator of transcription 5b (STAT5b) is constitutively activated in many human tumors. Activity of STAT5b requires binding of its Src homology 2 (SH2) domain to certain phosphotyrosine-containing sequences. We have developed a high-throughput assay based on fluorescence polarization that allows screening of chemical libraries for compounds that inhibit STAT5b by interfering with the function of its SH2 domain. The assay, which is based on binding between a fluorescein-labeled phosphotyrosine peptide derived from the erythropoietin receptor to the STAT5b SH2 domain, is stable with regard to dimethyl sulfoxide concentration and time and has a Z' value of 0.66+/-0.11 in a 384-well format.
信号转导及转录激活蛋白5b（STAT5b）在许多人类肿瘤中持续激活。STAT5b的活性需要其Src同源2（SH2）结构域与某些含磷酸酪氨酸的序列结合。我们开发了一种基于荧光偏振的高通量检测方法，可用于筛选化学文库，以寻找通过干扰其SH2结构域功能来抑制STAT5b的化合物。该检测方法基于源自促红细胞生成素受体的荧光素标记的磷酸酪氨酸肽与STAT5b SH2结构域之间的结合，在二甲基亚砜浓度和时间方面具有稳定性，在384孔板形式下的Z'值为0.66±0.11。
