Berg Thorsten, Cohen Steven B, Desharnais Joel, Sonderegger Corinna, Maslyar Daniel J, Goldberg Joel, Boger Dale L, Vogt Peter K
Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3830-5. doi: 10.1073/pnas.062036999. Epub 2002 Mar 12.
Myc is a transcriptional regulator of the basic helix-loop-helix leucine zipper protein family. It has strong oncogenic potential, mutated or virally transduced forms of Myc induce lymphoid tumors in animals, and deregulated expression of Myc is associated with numerous types of human cancers. For its oncogenic activity, Myc must dimerize with the ubiquitously expressed basic helix-loop-helix leucine zipper protein Max. This requirement for dimerization may allow control of Myc activity with small molecules that interfere with Myc/Max dimerization. We have measured Myc/Max dimerization with fluorescence resonance energy transfer and have screened combinatorial chemical libraries for inhibitors of dimerization. Candidate inhibitors were isolated from a peptidomimetics library. Inhibition of Myc/Max interaction was validated by ELISA and electrophoretic mobility-shift assay. Two of the candidate inhibitors also interfere with Myc-induced oncogenic transformation in chicken embryo fibroblast cultures. Our work provides proof of principle for the identification of small molecule inhibitors of protein-protein interactions by using high-throughput screens of combinatorial chemical libraries.
Myc是碱性螺旋-环-螺旋亮氨酸拉链蛋白家族的转录调节因子。它具有很强的致癌潜力，Myc的突变形式或病毒转导形式可在动物中诱发淋巴瘤，Myc的表达失调与多种人类癌症相关。就其致癌活性而言，Myc必须与普遍表达的碱性螺旋-环-螺旋亮氨酸拉链蛋白Max二聚化。这种对二聚化的需求可能使得能够用干扰Myc/Max二聚化的小分子来控制Myc的活性。我们用荧光共振能量转移测量了Myc/Max二聚化，并针对二聚化抑制剂筛选了组合化学文库。候选抑制剂是从拟肽文库中分离出来的。通过酶联免疫吸附测定和电泳迁移率变动分析验证了对Myc/Max相互作用的抑制作用。其中两种候选抑制剂也会干扰鸡胚成纤维细胞培养中Myc诱导的致癌转化。我们的工作为通过组合化学文库的高通量筛选来鉴定蛋白质-蛋白质相互作用的小分子抑制剂提供了原理证明。
