Screening for Inhibitors of Low-Affinity Epigenetic Peptide-Protein Interactions: An AlphaScreen™-Based Assay for Antagonists of Methyl-Lysine Binding Proteins
Screening for Inhibitors of Low-Affinity Epigenetic Peptide-Protein Interactions: An AlphaScreen™-Based Assay for Antagonists of Methyl-Lysine Binding Proteins
复制标题
DOI：
10.1177/1087057109352902
复制
发表时间：
2010-01-01
影响因子：
--
通讯作者：
Janzen, William P.
中科院分区：
文献类型：
--
作者：
Wigle, Tim J.;Herold, J. Martin;Janzen, William P.
组蛋白编码包括许多翻译后修饰，这些修饰主要发生在组蛋白尾肽中。这些标记的身份和位置由各种组蛋白结合蛋白读取，这些蛋白正在成为细胞分化和发育的重要调节因子，并越来越多地与多种疾病状态有关。作者描述了发现甲基赖氨酸结合蛋白抑制剂的第一个高通量筛选试验的发展，该试验将用于启动对这一广泛目标类别的全面发现工作。他们专注于开发一种基于Alphascreen T的恶性脑瘤(MBT)结构域蛋白的分析方法，这种蛋白与组蛋白尾肽中存在的赖氨酸残基的低甲基化状态结合。该方法利用Alphascreen T珠的亲和力，扫清了组蛋白结合蛋白与其同源多肽的微摩尔结合常数较低所带来的障碍。该分析也适用于其他甲基赖氨酸结合蛋白家族，并有可能用于筛选新的小分子，将其用作细胞重新编程和最终药物发现的研究工具。(生物分子筛选杂志2010：62-71)
The histone code comprises many posttranslational modifications that occur mainly in histone tail peptides. the identity and location of these marks are read by a variety of histone-binding proteins that are emerging as important regulators of cellular differentiation and development and are increasingly being implicated in numerous disease states. the authors describe the development of the first high-throughput screening assay for the discovery of inhibitors of methyl-lysine binding proteins that will be used to initiate a full-scale discovery effort for this broad target class. they focus on the development of an Alphascreen T-based assay for malignant brain tumor (MBT) domain-containing proteins, which bind to the lower methylation states of lysine residues present in histone tail peptides. this assay takes advantage of the avidity of the Alphascreen T beads to clear the hurdle to assay development presented by the low micromolar binding constants of the histone binding proteins for their cognate peptides. the assay is applicable to other families of methyl-lysine binding proteins, and it has the potential to be used in screening efforts toward the discovery of novel small molecules with utility as research tools for cellular reprogramming and ultimately drug discovery. (Journal of Biomolecular Screening 2010: 62-71)