Anil Burcu, Riedinger Christiane, Endicott Jane A, Noble Martin E M
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, England.
Acta Crystallogr D Biol Crystallogr. 2013 Aug;69(Pt 8):1358-66. doi: 10.1107/S0907444913004459. Epub 2013 Jul 13.
The p53-binding site of MDM2 holds great promise as a target for therapeutic intervention in MDM2-amplified p53 wild-type forms of cancer. Despite the extensive validation of this strategy, there are relatively few crystallographically determined co-complex structures for small-molecular inhibitors of the MDM2-p53 interaction available in the PDB. Here, a surface-entropy reduction mutant of the N-terminal domain of MDM2 that has been designed to enhance crystallogenesis is presented. This mutant has been validated by comparative ligand-binding studies using differential scanning fluorimetry and fluorescence polarization anisotropy and by cocrystallization with a peptide derived from p53. Using this mutant, the cocrystal structure of MDM2 with the benchmark inhibitor Nutlin-3a has been determined, revealing subtle differences from the previously described co-complex of MDM2 with Nutlin-2.
MDM2的p53结合位点作为MDM2扩增的p53野生型癌症治疗干预的靶点具有很大潜力。尽管该策略已得到广泛验证，但在蛋白质数据库（PDB）中，针对MDM2-p53相互作用的小分子抑制剂，通过晶体学确定的共复合物结构相对较少。在此，展示了一种为增强结晶作用而设计的MDM2 N端结构域的表面熵降低突变体。该突变体已通过差示扫描荧光法和荧光偏振各向异性进行的比较配体结合研究以及与源自p53的肽共结晶得到验证。利用该突变体，已确定MDM2与基准抑制剂Nutlin-3a的共晶体结构，揭示了与先前描述的MDM2与Nutlin-2的共复合物的细微差异。
