Detecting protein–protein interactions based on kinase-mediated growth induction of mammalian cells

• DOI: 10.1038/srep06127
• Publication Date: 2014-08-19T09:04:48Z
• Authors: Satoru Mabe; Teruyuki Nagamune; Masahiro Kawahara
• Keywords: 0301 basic medicine; 0303 health sciences; Proto-Oncogene Proteins c-mdm2; Article; Tacrolimus; 6. Clean water; Cell Line; 3. Good health; Tacrolimus Binding Proteins; Mice; Proto-Oncogene Proteins c-kit; 03 medical and health sciences; Amino Acid Substitution; Protein Interaction Mapping; Animals; Protein Interaction Domains and Motifs; Tumor Suppressor Protein p53; Peptides; Dimerization; Cell Proliferation

Detection of protein–protein interactions (PPIs) is important for understanding numerous processes in mammalian cells; however, existing PPI detection methods often give significant background signals. Here, we propose a novel PPI-detection method based on kinase-mediated growth induction cells. In this method, target proteins are fused to the intracellular domain c-kit (c-kit ICD) and expressed interleukin-3-dependent The induces dimerization activation ICDs, which leads cell absence interleukin-3. Using system, successfully detected ligand-dependent homo-interaction FKBPF36V hetero-interaction FKBP FRBT2098L, as well constitutive interaction between MDM2 known peptide inhibitor. Intriguingly, cells expressing high-affinity chimeras selected from mixture populations dominantly low-affinity chimeras. These results indicate that can detect PPIs with low levels suitable inhibitor screening.