Putative protein partners for the human CPI-17 protein revealed by bacterial two-hybrid screening

•Using two-hybrid system, several binding partners of CPI-17 were identified.•Plakoglobin was identified as a real binding partner of CPI-17 in human cells.•The phosphorylation of CPI-17 interrupted its interaction with plakoglobin.

We have previously demonstrated that PKC-potentiated inhibitory protein of protein phosphatase-1 (CPI-17) is expressed in lung endothelium. CPI-17, a specific inhibitor of myosin light chain phosphatase (MLCP), is involved in the endothelial cytoskeletal and barrier regulation. In this paper, we report the identification of fourteen putative CPI-17 interacting proteins in the lung using BacterioMatch Two-Hybrid System. Five of them: plectin 1 isoform 1, alpha II spectrin, OK/SW-CL.16, gelsolin isoform a, and junction plakoglobin are involved in actin cytoskeleton organization and cell adhesion, suggesting possible significance of these binding partners in CPI-17-mediated cytoskeletal reorganization of endothelial cells. Furthermore, we confirmed the specific interaction between plakoglobin and CPI-17, which is affected by the phosphorylation status of CPI-17 in human lung microvascular endothelial cells.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide