Rega Michele F, Reed John C, Pellecchia Maurizio
Burnham Institute for Medical Research, 10901 North Torrey Pines Rd., La Jolla, CA 92037, USA.
Bioorg Chem. 2007 Apr;35(2):113-20. doi: 10.1016/j.bioorg.2006.07.006. Epub 2006 Sep 22.
Anti-apoptotic Bcl-2-family proteins (Bcl-2, Bcl-x(L), Bfl-1, Mcl-1, Bcl-W and Bcl-B) have been recently validated as drug discovery targets for cancer, owed to their ability to confer tumor resistance to chemotherapy or radiation. The anti-apoptotic activity of Bcl-2 proteins is due to their ability to heterodimerize with their pro-apoptotic counterparts (proteins such as Bad, Bim or Bid) via a conserved peptide region termed BH3. Thus, molecules that mimic pro-apoptotic BH3 domains represent a direct approach to overcoming the protective effects of anti-apoptotic proteins such as Bcl-2 and Bcl-x(L). Here, we report on the development and evaluation of two novel Lanthanide-based assays that are formatted for high-throughput screening of small molecules capable of antagonizing BH3-Bcl-2 interactions. The assay conditions, robustness and reproducibility (Z' factors) are described. These assays represent useful tools to enable further studies in the search for novel, safe and effective anti-cancer agents targeting Bcl-2-family proteins.
抗凋亡的Bcl-2家族蛋白（Bcl-2、Bcl-x(L)、Bfl-1、Mcl-1、Bcl-W和Bcl-B）最近已被确认为癌症药物研发的靶点，这是因为它们能够赋予肿瘤对化疗或放疗的抗性。Bcl-2蛋白的抗凋亡活性归因于其能够通过一个称为BH3的保守肽区域与促凋亡对应蛋白（如Bad、Bim或Bid等蛋白）形成异二聚体。因此，模拟促凋亡BH3结构域的分子是克服抗凋亡蛋白（如Bcl-2和Bcl-x(L)）保护作用的直接方法。在此，我们报告了两种新型基于镧系元素的检测方法的开发和评估，这些方法被设计用于高通量筛选能够拮抗BH3-Bcl-2相互作用的小分子。文中描述了检测条件、稳健性和重现性（Z'因子）。这些检测方法是有用的工具，能够促进在寻找靶向Bcl-2家族蛋白的新型、安全且有效的抗癌药物方面的进一步研究。
