Institut Pasteur, Unité Récepteurs-Canaux, CNRS UMR 3571, Paris, France.
Institut Pasteur, Plate-forme de Biophysique Moléculaire, CNRS UMR 3528, Paris, France.
Methods Mol Biol. 2021;2256:89-124. doi: 10.1007/978-1-0716-1166-1_6.
PDZ domains are small globular domains involved in protein-protein interactions. They participate in a wide range of critical cellular processes. These domains, very abundant in the human proteome, are widely studied by high-throughput interactomics approaches and by biophysical and structural methods. However, the quality of the results is strongly related to the optimal folding and solubility of the domains. We provide here a detailed description of protocols for a strict quality assessment of the PDZ constructs. We describe appropriate experimental approaches that have been selected to overcome the small size of such domains to check the purity, identity, homogeneity, stability, and folding of samples.
PDZ 结构域是参与蛋白质-蛋白质相互作用的小型球形结构域。它们参与了广泛的关键细胞过程。这些结构域在人类蛋白质组中非常丰富,通过高通量相互作用组学方法以及生物物理和结构方法进行了广泛的研究。然而,结果的质量与结构域的最佳折叠和可溶性密切相关。我们在这里提供了详细的 PDZ 结构域构建体严格质量评估协议描述。我们描述了适当的实验方法,这些方法已被选中克服了这些结构域的小尺寸,以检查样品的纯度、身份、均一性、稳定性和折叠。