Sotelo Natalia S, Schepens Jan T G, Valiente Miguel, Hendriks Wiljan J A J, Pulido Rafael
Centro de Investigación Príncipe Felipe, Valencia, Spain.
Department of Cell Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands.
Methods. 2015 May;77-78:147-56. doi: 10.1016/j.ymeth.2014.10.017. Epub 2014 Oct 22.
Protein modular interactions mediated by PDZ domains are essential for the establishment of functional protein networks controlling diverse cellular functions. The tumor suppressor PTEN possesses a C-terminal PDZ-binding motif (PDZ-BM) that is recognized by a specific set of PDZ domains from scaffolding and regulatory proteins. Here, we review the current knowledge on PTEN-PDZ domain interactions and tumor suppressor networks, describe methodology suitable to analyze these interactions, and report the binding of PTEN and the PDZ domain-containing protein tyrosine phosphatase PTPN13. Yeast two-hybrid and GST pull-down analyses showed that PTEN binds to PDZ2/PTPN13 domain in a manner that depends on the specific PTPN13 PDZ domain arrangement involving the interdomain region between PDZ1 and PDZ2. Furthermore, a specific binding profile of PTEN to PDZ2/PTPN13 domain was observed by mutational analysis of the PTEN PDZ-BM. Our results disclose a PDZ-mediated physical interaction of PTEN and PTPN13 with potential relevance in tumor suppression and cell homeostasis.
由PDZ结构域介导的蛋白质模块化相互作用对于建立控制多种细胞功能的功能性蛋白质网络至关重要。肿瘤抑制因子PTEN具有一个C端PDZ结合基序（PDZ-BM），可被来自支架蛋白和调节蛋白的一组特定PDZ结构域识别。在这里，我们综述了关于PTEN-PDZ结构域相互作用和肿瘤抑制网络的现有知识，描述了适用于分析这些相互作用的方法，并报道了PTEN与含PDZ结构域的蛋白酪氨酸磷酸酶PTPN13的结合。酵母双杂交和GST下拉分析表明，PTEN以一种依赖于涉及PDZ1和PDZ2之间结构域间区域的特定PTPN13 PDZ结构域排列的方式与PDZ2/PTPN13结构域结合。此外，通过对PTEN PDZ-BM的突变分析观察到PTEN与PDZ2/PTPN13结构域的特异性结合谱。我们的结果揭示了PTEN和PTPN13之间由PDZ介导的物理相互作用，这在肿瘤抑制和细胞稳态中可能具有相关性。
