Inserm UMRS1131, Institut de Génétique Moléculaire, Université Paris 7, Hôpital St. Louis, F-75010 Paris, France.
Biochemistry-Molecular Biology, Faculty of Science, Universidad de la República, Iguá 4225, Montevideo 11400, Uruguay.
Biomolecules. 2020 Dec 30;11(1):36. doi: 10.3390/biom11010036.
The p53 and Mouse double minute 2 (MDM2) proteins are hubs in extensive networks of interactions with multiple partners and functions. Intrinsically disordered regions help to adopt function-specific structural conformations in response to ligand binding and post-translational modifications. Different techniques have been used to dissect interactions of the p53-MDM2 pathway, in vitro, in vivo, and in situ each having its own advantages and disadvantages. This review uses the p53-MDM2 to show how different techniques can be employed, illustrating how a combination of in vitro and in vivo techniques is highly recommended to study the spatio-temporal location and dynamics of interactions, and to address their regulation mechanisms and functions. By using well-established techniques in combination with more recent advances, it is possible to rapidly decipher complex mechanisms, such as the p53 regulatory pathway, and to demonstrate how protein and nucleotide ligands in combination with post-translational modifications, result in inter-allosteric and intra-allosteric interactions that govern the activity of the protein complexes and their specific roles in oncogenesis. This promotes elegant therapeutic strategies that exploit protein dynamics to target specific interactions.
p53 和 Mouse double minute 2（MDM2）蛋白是与多个伴侣和功能广泛相互作用网络的枢纽。无规卷曲区域有助于在配体结合和翻译后修饰的情况下采用特定功能的结构构象。已经使用了不同的技术来剖析 p53-MDM2 途径的相互作用，包括体外、体内和原位，每种方法都有其自身的优点和缺点。这篇综述使用 p53-MDM2 来展示如何使用不同的技术，说明如何结合使用体外和体内技术来研究相互作用的时空定位和动态，以及解决其调节机制和功能。通过结合使用成熟的技术和最近的进展，可以快速破译复杂的机制，例如 p53 调节途径，并证明蛋白质和核苷酸配体与翻译后修饰相结合如何导致变构相互作用和同型变构相互作用，从而控制蛋白复合物的活性及其在肿瘤发生中的特定作用。这促进了利用蛋白动力学来靶向特定相互作用的优雅治疗策略。
